JP2017153493A

JP2017153493A - Cell culture apparatus

Info

Publication number: JP2017153493A
Application number: JP2017118331A
Authority: JP
Inventors: 中川　博之; Hiroyuki Nakagawa; 博之中川; 誠太郎 ▲高▼城; Seitaro Takagi; 恒水野; Hisashi Mizuno; 敏行萩山; Toshiyuki Hagiyama; 篤軌吉村; Atsuki Yoshimura; 康彦荻巣; Yasuhiko Ogisu; 康彦篠田; Yasuhiko Shinoda; 賢田中; Ken Tanaka; 千香子佐藤; Chikako Sato
Original assignee: Yamagata University NUC; Toyoda Gosei Co Ltd
Current assignee: Yamagata University NUC; Toyoda Gosei Co Ltd
Priority date: 2017-06-16
Filing date: 2017-06-16
Publication date: 2017-09-07
Anticipated expiration: 2034-04-28
Also published as: JP6840629B2

Abstract

PROBLEM TO BE SOLVED: To hold a cell culture membrane with sufficiently high reliability while suppressing the requirement for dimensional accuracy of the constituent member.SOLUTION: A cell culture apparatus is equipped with a cylindrical first inner cylinder. The cell culture apparatus is also equipped with a cell culture membrane arranged so as to entirely cover one end of the first inner cylinder, the cell culture membrane being bent to the outside of the first inner cylinder along the periphery of the one end of the first inner cylinder. The cell culture apparatus is also equipped with an outer cylinder which is formed in cylinder form and houses interior the first inner cylinder so that the height direction of the outer cylinder corresponds to that of the first cylinder, wherein a fist slit extending toward the height direction is formed and the cell culture membrane is secured between the inner wall surface of the outer cylinder and the outer surface of the first inner cylinder, in a region where the outer cylinder is overlapped with at least the one end part of the first inner cylinder.SELECTED DRAWING: Figure 1

Description

本発明は、細胞培養器具に関するものである。 The present invention relates to a cell culture instrument.

細胞培養技術は、創薬、再生医療などの種々の分野で有用な技術として近年注目されている。足場依存性の細胞を培養する際には、一般に、細胞の足場となる基材である細胞培養用膜が用いられる。従来、このような細胞培養用膜を備える細胞培養器具として、例えば、互いに係合する凹凸形状を有する２つのリング状部材の間に細胞培養用膜を配置し、上記凹凸形状の部位において細胞培養用膜の外周を挟み込んで細胞培養用膜を保持する器具が提案されている（例えば、特許文献１参照）。 In recent years, cell culture techniques have attracted attention as useful techniques in various fields such as drug discovery and regenerative medicine. When culturing anchorage-dependent cells, a cell culture membrane that is a base material that becomes a scaffold for cells is generally used. Conventionally, as a cell culture instrument provided with such a cell culture membrane, for example, a cell culture membrane is disposed between two ring-shaped members having concave and convex shapes that engage with each other, and cell culture is performed at the concave and convex portions. An instrument that holds the cell culture membrane by sandwiching the outer periphery of the membrane is proposed (see, for example, Patent Document 1).

特開２００５−３４８７３６号公報JP 2005-348736 A 特表平１１−５１０３９４号公報Japanese National Patent Publication No. 11-510394

しかしながら、細胞培養用膜は、一般に数μｍ〜数十μｍ程度の薄さであり、このように薄い細胞培養用膜を確実に保持するためには、細胞培養用膜の外周を挟み込むための既述したリング状部材における凹凸形状の寸法精度を、十分に高くする必要があった。このように高い寸法精度が要求されると、細胞培養器具の製造工程の煩雑化やコストの増加を引き起こす可能性がある。そのため、細胞培養器具の構成部材の寸法精度に対する要求を抑えつつ、十分に高い信頼性で細胞培養用膜を保持できる細胞培養器具が望まれていた。 However, the cell culture membrane is generally about several μm to several tens of μm thin, and in order to securely hold such a thin cell culture membrane, the existing membrane for sandwiching the outer periphery of the cell culture membrane is used. It is necessary to sufficiently increase the dimensional accuracy of the uneven shape in the ring-shaped member described above. When such a high dimensional accuracy is required, there is a possibility that the manufacturing process of the cell culture instrument becomes complicated and the cost increases. Therefore, there has been a demand for a cell culture instrument that can hold the cell culture membrane with sufficiently high reliability while suppressing the demand for dimensional accuracy of the components of the cell culture instrument.

本発明は、上述の課題の少なくとも一部を解決するためになされたものであり、以下の形態として実現することが可能である。 SUMMARY An advantage of some aspects of the invention is to solve at least a part of the problems described above, and the invention can be implemented as the following forms.

（１）本発明の一形態によれば、細胞培養器具が提供される。この細胞培養器具は、筒状の第１の内筒と；前記第１の内筒の一方の端部全体を覆って配置された細胞培養用膜であって、前記第１の内筒の前記一方の端部の外周に沿って、前記第１の内筒の外側面側へと折り曲げられた細胞培養用膜と；筒状に形成されて、前記第１の内筒と高さ方向が一致するように前記第１の内筒を内部に収容する外筒であって、少なくとも前記第１の内筒の前記一方の端部と重なる領域において、前記高さ方向に延びる第１のスリットが形成されると共に、前記外筒の内壁面と前記第１の内筒の外側面との間で、前記細胞培養用膜を挟持する外筒と；を備える。
この形態の細胞培養器具によれば、第１の内筒の一方の端部と重なる領域において高さ方向に延びる第１のスリットを設けて、外筒の内壁面と第１の内筒の外側面との間で細胞培養用膜を挟持するため、各部材の寸法精度に対する要求を抑えつつ十分に高い信頼性で、細胞培養用膜を保持することが可能になる。その結果、細胞培養器具の製造工程の煩雑化やコストの増加を抑えることができる。 (1) According to one aspect of the present invention, a cell culture instrument is provided. The cell culture instrument is a cylindrical first inner cylinder; a membrane for cell culture arranged so as to cover the entire one end of the first inner cylinder, and the first inner cylinder A membrane for cell culture bent to the outer surface side of the first inner cylinder along the outer periphery of one end; formed in a cylindrical shape, the height direction of which coincides with the first inner cylinder As described above, an outer cylinder that accommodates the first inner cylinder therein, and a first slit that extends in the height direction is formed at least in a region that overlaps the one end of the first inner cylinder. And an outer cylinder that sandwiches the cell culture membrane between an inner wall surface of the outer cylinder and an outer surface of the first inner cylinder.
According to the cell culture instrument of this aspect, the first slit extending in the height direction is provided in a region overlapping with one end of the first inner cylinder, and the inner wall surface of the outer cylinder and the outer side of the first inner cylinder are provided. Since the cell culture membrane is sandwiched between the side surfaces, it is possible to hold the cell culture membrane with sufficiently high reliability while suppressing the demand for dimensional accuracy of each member. As a result, the complexity of the manufacturing process of the cell culture instrument and the increase in cost can be suppressed.

（２）上記形態の細胞培養器具において、前記第１のスリットは、前記高さ方向に沿って前記外筒全体に形成されていることとしてもよい。この形態の細胞培養器具によれば、一方の端部に細胞培養用膜を配置した第１の内筒を外筒に嵌め込む動作を容易化することができる。 (2) The cell culture instrument of the said form WHEREIN: A said 1st slit is good also as being formed in the said outer cylinder whole along the said height direction. According to the cell culture instrument of this form, the operation | movement which fits the 1st inner cylinder which has arrange | positioned the film | membrane for cell culture in one edge part in an outer cylinder can be facilitated.

（３）上記形態の細胞培養器具において、前記外筒は、前記第１の内筒の両端部のうちの前記一方の端部により近い側の端部において、該端部の開口を跨ぐように該開口の外周の異なる位置間を連結した第１の把持部を備えることとしてもよい。
このような構成とすれば、把持部を把持することによって細胞培養器具を吊り下げることが可能になり、細胞培養器具を取り扱う操作を容易化することができる。 (3) In the cell culture instrument of the above aspect, the outer cylinder may straddle the opening of the end at the end closer to the one end of the both ends of the first inner cylinder. It is good also as providing the 1st holding part which connected between different positions of the perimeter of this opening.
With such a configuration, it becomes possible to suspend the cell culture instrument by gripping the grip portion, and the operation for handling the cell culture instrument can be facilitated.

（４）上記形態の細胞培養器具において、さらに、前記第１の内筒と高さ方向が同じである筒状に形成されて、前記第１の内筒の前記一方の端部側で前記細胞培養用膜を介して前記第１の内筒と接するように前記外筒内に収容される第２の内筒を備えることとしてもよい。
このような構成とすれば、第１の内筒の外側に面する細胞培養用膜の面上に細胞を播種する際に、第１のスリットを介した細胞培養器具の外部への細胞の流出が第２の内筒によって抑えられる。そのため、播種する細胞数の精度を向上させることができる。 (4) In the cell culture instrument of the above aspect, the cell is further formed in a cylindrical shape having the same height direction as the first inner cylinder, and the cell on the one end side of the first inner cylinder It is good also as providing the 2nd inner cylinder accommodated in the said outer cylinder so that the said 1st inner cylinder may be contacted via the film | membrane for culture | cultivation.
With such a configuration, when cells are seeded on the surface of the cell culture membrane facing the outside of the first inner cylinder, the outflow of the cells to the outside of the cell culture instrument through the first slit. Is suppressed by the second inner cylinder. Therefore, the accuracy of the number of cells to be seeded can be improved.

（５）上記形態の細胞培養器具において、前記第２の内筒は、前記高さ方向の力を加えることにより、前記外筒内で前記高さ方向にスライド可能であって、スライドさせた位置にて保持可能であることとしてもよい。
このような構成とすれば、第２の内筒内で露出する細胞培養用膜上に細胞を播種した後に、第２の内筒をスライドさせて細胞培養用膜から離間させ、さらに細胞培養器具を上下反転させることにより、細胞培養用膜における上記細胞を播種した面上に残留するエアを、第１のスリットを介して容易に排出させることが可能になる。 (5) In the cell culture instrument of the above aspect, the second inner cylinder is slidable in the height direction in the outer cylinder by applying a force in the height direction, and is slid It is good also as being able to hold at.
With such a configuration, after seeding the cells on the cell culture membrane exposed in the second inner cylinder, the second inner cylinder is slid to be separated from the cell culture membrane. , The air remaining on the surface of the cell culture membrane seeded with the cells can be easily discharged through the first slit.

（６）上記形態の細胞培養器具において、前記外筒に形成される前記第１のスリットは、前記第１の内筒の前記一方の端部と前記外筒とが重なる位置から、前記第１の内筒から高さ方向に離間する方向に向かって延びるように形成されていることとしてもよい。
このような構成とすれば、前記第１の内筒の外部に面する前記細胞培養用膜の面上に細胞を播種した後に、細胞培養器具を上下反転させたときに、細胞培養用膜における上記細胞を播種した面上に残留するエアを、第１のスリットを介して容易に排出させることが可能になる。 (6) In the cell culture instrument of the above aspect, the first slit formed in the outer cylinder is located at a position where the one end of the first inner cylinder and the outer cylinder overlap with each other. It is good also as forming so that it may extend toward the direction spaced apart from the inner cylinder of a height direction.
With such a configuration, when cells are seeded on the surface of the cell culture membrane facing the outside of the first inner cylinder and then the cell culture instrument is turned upside down, the cell culture membrane The air remaining on the surface seeded with the cells can be easily discharged through the first slit.

（７）上記形態の細胞培養器具において、前記第１の内筒は、該第１の内筒の両端部のうちの他方の端部において、該他方の端部の開口を跨ぐように該他方の端部の開口の外周の異なる位置間を連結した第２の把持部を備えることとしてもよい。
このような構成とすれば、第１の内筒の端部のうち、前記他方の端部側が鉛直方向上方となる向きに細胞培養器具が配置されるときに、第２の把持部を把持することによって細胞培養器具を吊り下げることが可能になり、細胞培養器具を取り扱う操作を容易化することができる。 (7) In the cell culture instrument of the above aspect, the first inner cylinder may be configured so that the other end of the first inner cylinder extends across the opening of the other end at the other end. It is good also as providing the 2nd holding | grip part which connected between the positions where the outer periphery of opening of the edge part differs.
With such a configuration, when the cell culture instrument is arranged in such a direction that the other end side of the end portion of the first inner cylinder is vertically upward, the second gripping portion is gripped. As a result, the cell culture instrument can be suspended and the operation of handling the cell culture instrument can be facilitated.

（８）上記形態の細胞培養器具において、前記第１の把持部は、前記第１の内筒の軸線上付近で交わる２本の線分状に形成されていることとしてもよい。
このような構成とすれば、第１又は第２の把持部を把持して細胞培養器具を吊り下げる際に、細胞培養用膜の水平を安定して保つことが容易となり、細胞培養用膜上で培養される細胞に対する望ましくない外的刺激（ノイズ）等を抑えることができる。 (8) In the cell culture instrument of the above aspect, the first grip portion may be formed in two line segments that intersect in the vicinity of the axis of the first inner cylinder.
With such a configuration, when the cell culture instrument is suspended by holding the first or second holding portion, it becomes easy to stably keep the cell culture membrane horizontal. Undesirable external stimuli (noise) and the like on the cells cultured in can be suppressed.

（９）上記形態の細胞培養器具において、前記第２の把持部は、前記第１の内筒の軸線上付近で交わる２本の線分状に形成されていることとしてもよい。
このような構成とすれば、第１又は第２の把持部を把持して細胞培養器具を吊り下げる際に、細胞培養用膜の水平を安定して保つことが容易となり、細胞培養用膜上で培養される細胞に対する望ましくない外的刺激（ノイズ）等を抑えることができる。 (9) In the cell culture instrument of the above aspect, the second grip portion may be formed in two line segments that intersect in the vicinity of the axis of the first inner cylinder.
With such a configuration, when the cell culture instrument is suspended by holding the first or second holding portion, it becomes easy to stably keep the cell culture membrane horizontal. Undesirable external stimuli (noise) and the like on the cells cultured in can be suppressed.

（１０）上記形態の細胞培養器具において、前記第１の内筒には；前記外筒に設けた前記第１のスリットと重なる位置において、前記一方の端部から前記高さ方向に延びる第２のスリットと；前記外筒に設けた前記第１のスリットと重なる位置であって、前記第１の内筒の前記一方の端部における前記第２のスリットが開口する位置において、前記第１の内筒の径方向外側に向かって前記第１のスリットを経由して前記外筒の外側に延びる一対の膜支持部と；が設けられていることとしてもよい。
このような構成とすれば、前記第１の内筒の外部に面する前記細胞培養用膜の面上に細胞を播種する際に、細胞培養用膜における細胞を播種する面の裏面に残留するエアを、第１のスリットおよび第２のスリットを介して排出させることが可能になる。 (10) In the cell culture instrument of the above aspect, the first inner cylinder; a second extending in the height direction from the one end portion at a position overlapping the first slit provided in the outer cylinder. The first slit provided in the outer cylinder and the first slit at the position where the second slit is opened at the one end of the first inner cylinder. And a pair of membrane support portions extending outward of the outer cylinder through the first slit toward the radially outer side of the inner cylinder.
With such a configuration, when cells are seeded on the surface of the cell culture membrane facing the outside of the first inner cylinder, the cells remain on the back surface of the surface of the cell culture membrane on which the cells are seeded. Air can be discharged through the first slit and the second slit.

（１１）上記形態の細胞培養器具において、前記第１の内筒は、該第１の内筒における前記一方の端部近傍の外側面であって、前記細胞培養用膜と接する位置に、凹凸形状が設けられていることとしてもよい。
このような構成とすれば、第１の内筒および外筒によって細胞培養用膜を挟持する際の保持力を高めることができる。 (11) In the cell culture instrument of the above aspect, the first inner cylinder is an uneven surface at an outer surface of the first inner cylinder in the vicinity of the one end and in contact with the cell culture membrane. A shape may be provided.
With such a configuration, it is possible to increase the holding force when the cell culture membrane is sandwiched between the first inner cylinder and the outer cylinder.

本発明は、装置以外の種々の形態で実現することも可能である。例えば、細胞培養器具用の外筒、細胞培養器具の製造方法、細胞培養用膜の保持方法、あるいは細胞培養器具を用いた細胞培養方法等の形態で実現することが可能である。 The present invention can be realized in various forms other than the apparatus. For example, it can be realized in the form of an outer tube for a cell culture instrument, a method for manufacturing a cell culture instrument, a method for holding a cell culture membrane, or a cell culture method using a cell culture instrument.

本発明の細胞培養器具によれば、第１の内筒の一方の端部と重なる領域において高さ方向に延びる第１のスリットを設けて、外筒の内壁面と第１の内筒の外側面との間で細胞培養用膜を挟持するため、各部材の寸法精度に対する要求を抑えつつ十分に高い信頼性で、細胞培養用膜を保持することが可能になる。その結果、細胞培養器具の製造工程の煩雑化やコストの増加を抑えることができる。 According to the cell culture instrument of the present invention, the first slit extending in the height direction is provided in a region overlapping with one end portion of the first inner cylinder, and the inner wall surface of the outer cylinder and the outer side of the first inner cylinder are provided. Since the cell culture membrane is sandwiched between the side surfaces, it is possible to hold the cell culture membrane with sufficiently high reliability while suppressing the demand for dimensional accuracy of each member. As a result, the complexity of the manufacturing process of the cell culture instrument and the increase in cost can be suppressed.

細胞培養器具の構成を表わす断面模式図である。It is a cross-sectional schematic diagram showing the structure of a cell culture instrument. 細胞培養器具の組み立ての様子を説明するための分解斜視図である。It is a disassembled perspective view for demonstrating the mode of an assembly of a cell culture instrument. 細胞培養器具の上面図である。It is a top view of a cell culture instrument. 細胞培養器具の構成を表わす断面模式図である。It is a cross-sectional schematic diagram showing the structure of a cell culture instrument. 細胞培養器具の組み立ての様子を説明するための分解斜視図である。It is a disassembled perspective view for demonstrating the mode of an assembly of a cell culture instrument. 細胞培養器具の構成を表わす断面模式図である。It is a cross-sectional schematic diagram showing the structure of a cell culture instrument. 第１の内筒の一部を拡大して示す斜視図である。It is a perspective view which expands and shows a part of 1st inner cylinder. 細胞培養器具の構成を表わす断面模式図である。It is a cross-sectional schematic diagram showing the structure of a cell culture instrument. 細胞培養器具の構成を表わす断面模式図である。It is a cross-sectional schematic diagram showing the structure of a cell culture instrument.

Ａ．第１の実施形態：
図１は、本発明の第１の実施形態である細胞培養器具１０の構成を表わす断面模式図であり、図２は、細胞培養器具１０の組み立ての様子を説明するための分解斜視図であり、図３は、細胞培養器具１０の上面図である。なお、図３では、図１に示す断面の位置をＡ−Ａ断面として示している。細胞培養器具１０は、外筒１２と、第１の内筒１４と、細胞培養用膜１６と、を備える。 A. First embodiment:
FIG. 1 is a schematic cross-sectional view showing a configuration of a cell culture instrument 10 according to the first embodiment of the present invention, and FIG. 2 is an exploded perspective view for explaining an assembly state of the cell culture instrument 10. FIG. 3 is a top view of the cell culture instrument 10. In addition, in FIG. 3, the position of the cross section shown in FIG. 1 is shown as AA cross section. The cell culture instrument 10 includes an outer cylinder 12, a first inner cylinder 14, and a cell culture membrane 16.

外筒１２は、筒状の部材であって、一方の端部１２ｏから他方の端部１２ｔまで高さ方向に延びる第１のスリット１２ｓが形成されている。外筒１２は、一方の端部１２ｏにおいて、この一方の端部１２ｏの開口を跨ぐように、開口の外周の異なる位置間を連結した第１の把持部１２ｈを備える。具体的には、第１の把持部１２ｈは、第１の内筒１４の軸線上で交わる２本の線分状、すなわち、上面視が十字形状となるように形成されている（図２および図３参照）。外筒１２は、例えば、樹脂材料やゴムにより形成することができる。外筒１２は、高さ方向に第１のスリット１２ｓを設けることにより、径方向に弾性変形可能となる材料により形成すればよい。また、外筒１２は、後述するように第１の内筒１４との間で細胞培養用膜１６を挟んで保持するため、径方向外側に弾性変形して第１の内筒１４と共に細胞培養用膜１６を挟持したときに、径方向内側に向かう十分な把持力を生じ得る材料により形成すればよい。 The outer cylinder 12 is a cylindrical member, and is formed with a first slit 12s extending in the height direction from one end 12o to the other end 12t. The outer cylinder 12 includes a first grip portion 12h that connects between different positions on the outer periphery of the opening so as to straddle the opening of the one end portion 12o at one end portion 12o. Specifically, the first grip portion 12h is formed so as to have two line segments that intersect on the axis of the first inner cylinder 14, that is, a cross shape when viewed from above (see FIG. 2 and FIG. 2). (See FIG. 3). The outer cylinder 12 can be formed of, for example, a resin material or rubber. The outer cylinder 12 may be formed of a material that can be elastically deformed in the radial direction by providing the first slit 12s in the height direction. Further, since the outer cylinder 12 holds the cell culture membrane 16 with the first inner cylinder 14 as will be described later, the outer cylinder 12 is elastically deformed radially outward and cell culture is performed together with the first inner cylinder 14. What is necessary is just to form with the material which can produce sufficient gripping force which goes to radial direction inner side when the film | membrane 16 is clamped.

第１の内筒１４は、筒状の部材であって、外筒１２よりも高さが小さく形成されている。第１の内筒１４の一方の端部１４ｏには、細胞培養用膜１６が配置され、他方の端部１４ｔには、外筒１２に設けた第１の把持部１２ｈと同様の第２の把持部１４ｈが設けられている。第１の内筒１４は、例えば、樹脂材料やガラスにより形成することができる。第１の内筒１４は、後述するように外筒１２との間で細胞培養用膜１６を挟んで保持するため、外筒１２の内壁面から径方向内側に向かう力が加えられたときに、実質的に変形しない剛性を実現できる材料により形成すればよい。 The first inner cylinder 14 is a cylindrical member and is formed with a height smaller than that of the outer cylinder 12. A cell culture membrane 16 is disposed on one end portion 14o of the first inner cylinder 14, and a second end similar to the first gripping portion 12h provided on the outer cylinder 12 is disposed on the other end portion 14t. A gripping portion 14h is provided. The first inner cylinder 14 can be formed of, for example, a resin material or glass. Since the first inner cylinder 14 holds the cell culture membrane 16 with the outer cylinder 12 as will be described later, a force directed radially inward from the inner wall surface of the outer cylinder 12 is applied. What is necessary is just to form with the material which can implement | achieve the rigidity which does not deform | transform substantially.

なお、後述するように、細胞培養器具１０は、細胞培養を行なう際には培養液中に浸漬して用いるため、外筒１２および第１の内筒１４は、培養細胞に影響し得る金属イオン等の成分が培養液中に実質的に溶出しない材料により形成すればよい。また、外筒１２および第１の内筒１４は、細胞培養器具１０を培養液中で浮遊させない比重を有する材料により形成することが望ましい。外筒１２および第１の内筒１４を樹脂により形成する場合には、例えば、ポリエチレン、ポリプロピレン（ＰＰ）、シリコーン樹脂、ポリテトラフルオロエチレン（ＰＴＦＥ）、ポリエチレンテレフタレート（ＰＥＴ）、およびポリメチルメタクリレート（ＰＭＭＡ）から選択される樹脂を用いることができる。 As will be described later, since the cell culture instrument 10 is used by immersing it in a culture solution when performing cell culture, the outer cylinder 12 and the first inner cylinder 14 are metal ions that can affect the cultured cells. Such a component may be formed of a material that does not substantially elute into the culture solution. The outer cylinder 12 and the first inner cylinder 14 are preferably formed of a material having a specific gravity that does not cause the cell culture instrument 10 to float in the culture solution. When the outer cylinder 12 and the first inner cylinder 14 are formed of resin, for example, polyethylene, polypropylene (PP), silicone resin, polytetrafluoroethylene (PTFE), polyethylene terephthalate (PET), and polymethyl methacrylate ( A resin selected from PMMA) can be used.

細胞培養用膜１６は、細胞培養を行なう際に培養細胞の足場となる膜である。細胞培養用膜１６としては、例えば、ポリウレタン膜、セルロース膜、コラーゲン膜、羊膜、ヒアルロン酸膜等から選択される膜を用いることができる。ここで、本実施形態では、細胞培養用膜１６を折り曲げて外筒１２と第１の内筒１４との間で挟持して細胞培養器具１０を用いるため、細胞培養用膜１６は、上記折り曲げおよび挟持に耐える弾性および強度を有すればよい。また、細胞培養用膜１６として、膜の厚み方向に貫通する多数の細孔を有する多孔質膜を用いる場合には、後述するように両面で異なる種類の細胞を培養して、異種細胞間の相互作用を解析することが可能になる。そのため、膜を厚み方向に貫通する貫通孔の形成が容易であり、弾性および強度が優れているという観点から、細胞培養用膜１６としては、特に多孔質ポリウレタン膜を用いることが望ましい。 The cell culture membrane 16 is a membrane that serves as a scaffold for cultured cells when cell culture is performed. As the cell culture membrane 16, for example, a membrane selected from a polyurethane membrane, a cellulose membrane, a collagen membrane, an amniotic membrane, a hyaluronic acid membrane and the like can be used. Here, in this embodiment, since the cell culture membrane 16 is bent and sandwiched between the outer cylinder 12 and the first inner cylinder 14 and the cell culture instrument 10 is used, the cell culture membrane 16 is bent as described above. It is only necessary to have elasticity and strength to withstand clamping. When a porous membrane having a large number of pores penetrating in the thickness direction of the membrane is used as the cell culture membrane 16, different types of cells are cultured on both sides as described later, It becomes possible to analyze the interaction. Therefore, it is desirable to use a porous polyurethane film as the cell culture film 16 from the viewpoint that it is easy to form a through-hole penetrating the film in the thickness direction and that the elasticity and strength are excellent.

細胞培養用膜１６として用いる多孔質ポリウレタン膜は、例えば以下のようにして製造することができる。まず、ポリエーテルポリオール等のポリオールと、芳香族イソシアネート等のイソシアネートと、希釈溶剤とを含む未硬化ポリウレタン原料を用意する。その後、基板上に、用意した未硬化ポリウレタン原料の層を形成する。そして、未硬化ポリウレタン原料の層に対して水蒸気を供給しながらポリウレタン原料の層を硬化させることにより、ポリウレタン多孔質膜が得られる。 The porous polyurethane membrane used as the cell culture membrane 16 can be produced, for example, as follows. First, an uncured polyurethane raw material containing a polyol such as a polyether polyol, an isocyanate such as an aromatic isocyanate, and a diluent solvent is prepared. Thereafter, a layer of the prepared uncured polyurethane raw material is formed on the substrate. A polyurethane porous membrane is obtained by curing the polyurethane raw material layer while supplying water vapor to the uncured polyurethane raw material layer.

未硬化ポリウレタン原料の層に対して水蒸気を供給する具体的な方法としては、例えば、水を入れた密閉容器内において、基板上の未硬化ポリウレタン原料の層の露出面が水に対向するように、未硬化ポリウレタン原料の層を配置する。そして、密閉容器内を所定の温度に維持して飽和蒸気圧にする。これにより、未硬化ポリウレタン原料の層の表面から水蒸気が供給されて、ポリウレタン原料中のイソシアネートと水蒸気とが反応して二酸化炭素を生じる。また、ポリオールとイソシアネートとによって硬化反応が進行する。このように、硬化途中のポリウレタンが発泡することにより、多孔質ポリウレタン膜が得られる。なお、上記した水蒸気の供給を伴う硬化反応における反応温度、反応時間、硬化時に供給する水蒸気の量、およびポリウレタン原料の組成から選択される条件を調節することにより、ポリウレタン多孔質膜の形状を制御することが可能になる。 As a specific method of supplying water vapor to the uncured polyurethane raw material layer, for example, in an airtight container containing water, the exposed surface of the uncured polyurethane raw material layer on the substrate faces the water. A layer of uncured polyurethane raw material is disposed. Then, the inside of the sealed container is maintained at a predetermined temperature to obtain a saturated vapor pressure. Thereby, water vapor is supplied from the surface of the layer of the uncured polyurethane raw material, and the isocyanate and the water vapor in the polyurethane raw material react to generate carbon dioxide. Further, the curing reaction proceeds with the polyol and the isocyanate. Thus, a porous polyurethane film is obtained by foaming the polyurethane in the middle of curing. The shape of the polyurethane porous membrane is controlled by adjusting the conditions selected from the reaction temperature, reaction time, amount of water vapor supplied at the time of curing, and the composition of the polyurethane raw material in the curing reaction involving the supply of water vapor as described above. It becomes possible to do.

外筒１２と第１の内筒１４と細胞培養用膜１６とを用いた細胞培養器具１０の組み立て方法は、図２に示すとおりである。すなわち、細胞培養器具１０を組み立てる際には、まず、第１の内筒１４の一方の端部１４ｏ（第２の把持部１４ｈを有しない側の端部）上に、細胞培養用膜１６を配置する。なお、図２では、細胞培養用膜１６を四角形状に表わしているが、異なる形状としてもよい。第１の内筒１４の端部（横断面）よりも十分に大きく、第１の内筒１４の一方の端部１４ｏ全体を覆うことが可能であって、外筒１２と第１の内筒１４とによって細胞培養用膜１６の周縁部を挟持したときに、十分な信頼性で細胞培養用膜１６を保持可能となる大きさであればよい。 A method for assembling the cell culture instrument 10 using the outer cylinder 12, the first inner cylinder 14, and the cell culture membrane 16 is as shown in FIG. That is, when the cell culture instrument 10 is assembled, first, the cell culture membrane 16 is placed on one end portion 14o (the end portion on the side not having the second gripping portion 14h) of the first inner cylinder 14. Deploy. In FIG. 2, the cell culture membrane 16 is shown in a quadrangular shape, but it may have a different shape. It is sufficiently larger than the end portion (transverse section) of the first inner cylinder 14 and can cover the entire one end portion 14o of the first inner cylinder 14, and the outer cylinder 12 and the first inner cylinder When the peripheral portion of the cell culture membrane 16 is sandwiched between the cell culture membrane 16 and the cell culture membrane 16, the cell culture membrane 16 may be of a size that can be held with sufficient reliability.

その後、第１の内筒１４に対して、一方の端部１４ｏ側から、外筒１２の他方の端部１２ｔ（第１の把持部１２ｈを有しない側の端部）を嵌め込む。これにより、細胞培養用膜１６は、第１の内筒１４の一方の端部１４ｏの外周に沿って折り曲げられ、折り曲げられた位置よりも外周側である周縁部は、第１の内筒１４の外側面と外筒１２の内壁面によって挟持されるようになる。本実施形態では、外筒１２の上記他方の端部１２ｔと、第１の内筒１４の他方の端部１４ｔ（第２の把持部１４ｈを有する側の端部）とが重なるまで、外筒１２を嵌め込む（図１（Ａ）参照）。 Thereafter, the other end portion 12t of the outer tube 12 (the end portion on the side not having the first gripping portion 12h) is fitted into the first inner tube 14 from the one end portion 14o side. As a result, the cell culture membrane 16 is bent along the outer periphery of one end portion 14o of the first inner cylinder 14, and the peripheral portion on the outer peripheral side of the bent position is the first inner cylinder 14. Between the outer surface of the outer cylinder 12 and the inner wall surface of the outer cylinder 12. In the present embodiment, the outer cylinder 12 until the other end 12t of the outer cylinder 12 and the other end 14t of the first inner cylinder 14 (the end on the side having the second gripping portion 14h) overlap. 12 is inserted (see FIG. 1A).

外筒１２の内径は、上記嵌め込みを行なう前は、第１の内筒１４の外径よりも小さく形成されている。そのため、外筒１２内に第１の内筒１４および細胞培養用膜１６を収納するために、第１の内筒１４の外側に外筒１２を嵌め込む際には、第１のスリット１２ｓが広がるように外筒１２が押し広げられて、外筒１２は、径方向外側に弾性変形する。これにより、外筒１２から第１の内筒１４側へと、径方向内側に向かう把持力が生じると共に、外筒１２と第１の内筒１４とが、細胞培養用膜１６を介して隙間無く接する。その結果、細胞培養用膜１６が、外筒１２と第１の内筒１４との間で保持される。なおここで、外筒１２の嵌め込みの際には、外筒１２の内径を、内筒１４の外径と略等しく、または若干大きくなるように予め拡径した状態で、内筒１４の外側に外筒１２を嵌め込み、その後、外筒１２をその弾性復元力により縮径させて、細胞培養膜１６を保持させてもよい。上記のように第１の内筒１４の外側に嵌め込まれた外筒１２は、後述するように細胞培養器具１０を上下反転させても外筒１２内における第１の内筒１４の相対的な位置が変化しない程度の把持力で、内筒１４を把持している。 The inner diameter of the outer cylinder 12 is formed smaller than the outer diameter of the first inner cylinder 14 before the fitting. Therefore, when the outer cylinder 12 is fitted outside the first inner cylinder 14 in order to accommodate the first inner cylinder 14 and the cell culture membrane 16 in the outer cylinder 12, the first slit 12 s is formed. The outer cylinder 12 is pushed and expanded so as to expand, and the outer cylinder 12 is elastically deformed radially outward. As a result, a gripping force directed radially inward from the outer cylinder 12 toward the first inner cylinder 14 is generated, and the outer cylinder 12 and the first inner cylinder 14 are spaced from each other via the cell culture membrane 16. Contact without any exception. As a result, the cell culture membrane 16 is held between the outer cylinder 12 and the first inner cylinder 14. Here, when the outer cylinder 12 is fitted, the inner diameter of the outer cylinder 12 is expanded outside in advance so that the inner diameter of the outer cylinder 12 is substantially equal to or slightly larger than the outer diameter of the inner cylinder 14. The outer tube 12 may be fitted, and then the outer tube 12 may be reduced in diameter by its elastic restoring force to hold the cell culture membrane 16. As described above, the outer cylinder 12 fitted to the outside of the first inner cylinder 14 is relative to the first inner cylinder 14 in the outer cylinder 12 even when the cell culture instrument 10 is turned upside down as will be described later. The inner cylinder 14 is gripped with a gripping force that does not change the position.

組み立てた細胞培養器具１０を細胞培養に用いる際には、細胞培養器具１０を、さらに細胞培養用のプレートのウエル１１内に入れて（図２参照）、ウエル１１内を培養液で満たせばよい。 When the assembled cell culture instrument 10 is used for cell culture, the cell culture instrument 10 is further placed in a well 11 of a cell culture plate (see FIG. 2), and the well 11 is filled with a culture solution. .

図１（Ａ）は、細胞培養用膜１６の一方の面（第１の内筒１４の外部に露出する面）上に細胞αを播種した様子を模式的に表わす。なお、図１では、培養液および細胞培養用プレートのウエル１１については記載を省略している。細胞培養用膜１６の一方の面上に細胞αを播種するには、外筒１２の一方の端部１２ｏが鉛直方向上方に位置するように細胞培養器具１０を培養液中に浸漬した状態で、細胞培養用膜１６の上記一方の面上に形成された外筒１２内の空間に対して、所定量の細胞αを供給すればよい。 FIG. 1A schematically shows a state in which cells α are seeded on one surface of the cell culture membrane 16 (a surface exposed to the outside of the first inner cylinder 14). In FIG. 1, the description of the culture solution and the well 11 of the cell culture plate is omitted. In order to seed cells α on one surface of the cell culture membrane 16, the cell culture instrument 10 is immersed in the culture solution so that one end 12 o of the outer cylinder 12 is positioned vertically upward. A predetermined amount of cells α may be supplied to the space in the outer cylinder 12 formed on the one surface of the cell culture membrane 16.

図１（Ｂ）は、細胞培養用膜１６の他方の面（第１の内筒１４の内部に露出する面）上に、細胞αとは異なる細胞βをさらに播種した様子を示す。細胞βを播種する際には、図１（Ａ）のように細胞αを播種して所定期間培養した後に、細胞培養器具１０を上下反転させ、細胞培養用膜１６の上記他方の面上に形成された外筒１２および第１の内筒１４内の空間に対して、所定量の細胞βを供給すればよい。細胞培養用膜１６として、厚み方向に貫通する多数の貫通孔を有する多孔質な膜を用いることで、上記のようにして異なる細胞を各々の面で培養し、異種細胞間の相互作用を解析することが可能になる。 FIG. 1B shows a state where cells β different from the cells α are further seeded on the other surface of the cell culture membrane 16 (surface exposed inside the first inner cylinder 14). When seeding the cells β, after seeding the cells α and culturing them for a predetermined period as shown in FIG. 1A, the cell culture instrument 10 is turned upside down and placed on the other surface of the cell culture membrane 16. A predetermined amount of cells β may be supplied to the space inside the formed outer cylinder 12 and first inner cylinder 14. By using a porous membrane having a large number of through-holes penetrating in the thickness direction as the membrane for cell culture 16, different cells are cultured on each side as described above, and the interaction between different types of cells is analyzed. It becomes possible to do.

以上のように構成された本実施形態の細胞培養器具１０によれば、高さ方向に延びる第１のスリット１２ｓを有する外筒１２を、細胞培養用膜１６を配置した側から第１の内筒１４の外側に嵌め込むため、外筒１２から第１の内筒１４側へと径方向内側に向かう把持力を生じさせて、外筒１２と第１の内筒１４とによって細胞培養用膜１６を挟持することができる。そのため、各部材の寸法精度に対する要求を抑えつつ十分に高い信頼性で、細胞培養用膜１６を保持することが可能になり、その結果、細胞培養器具の製造工程の煩雑化やコストの増加を抑えることができる。 According to the cell culture instrument 10 of the present embodiment configured as described above, the outer cylinder 12 having the first slit 12s extending in the height direction is connected to the first inner side from the side where the cell culture membrane 16 is disposed. In order to be fitted to the outside of the cylinder 14, a gripping force directed radially inward from the outer cylinder 12 toward the first inner cylinder 14 is generated, and the cell culture membrane is formed by the outer cylinder 12 and the first inner cylinder 14. 16 can be clamped. Therefore, it becomes possible to hold the cell culture membrane 16 with sufficiently high reliability while suppressing the demand for the dimensional accuracy of each member. As a result, the manufacturing process of the cell culture instrument is complicated and the cost is increased. Can be suppressed.

また、本実施形態の細胞培養器具１０によれば、細胞培養器具１０の組み立て動作を簡素化することができる。すなわち、第１の内筒１４の一方の端部１４ｏ上に細胞培養用膜１６を配置して、第１の内筒１４の一方の端部１４ｏ側の開口に沿って細胞培養用膜１６を折り曲げつつ、第１の内筒１４および細胞培養用膜１６の外側に外筒１２を嵌め込むという簡便な動作により、細胞培養器具１０を組み立てることができる。 Moreover, according to the cell culture instrument 10 of this embodiment, the assembly operation | movement of the cell culture instrument 10 can be simplified. That is, the cell culture membrane 16 is disposed on one end portion 14o of the first inner cylinder 14, and the cell culture membrane 16 is disposed along the opening on the one end portion 14o side of the first inner cylinder 14. The cell culture instrument 10 can be assembled by a simple operation of fitting the outer cylinder 12 outside the first inner cylinder 14 and the cell culture membrane 16 while being bent.

特に、本実施形態の細胞培養器具１０によれば、外筒１２に形成される第１のスリット１２ｓが、外筒１２の高さ方向全体にわたって形成されているため、端部に細胞培養用膜１６を配置した第１の内筒１４を外筒１２に嵌め込む動作を容易化することができる。 In particular, according to the cell culture instrument 10 of the present embodiment, since the first slit 12s formed in the outer cylinder 12 is formed over the entire height direction of the outer cylinder 12, a cell culture membrane is formed at the end. The operation | movement which fits the 1st inner cylinder 14 which has arrange | positioned 16 in the outer cylinder 12 can be made easy.

また、本実施形態の細胞培養器具１０によれば、外筒１２の第１の把持部１２ｈおよび第１の内筒１４の第２の把持部１４ｈとして、外筒１２あるいは第１の内筒１４の端部の開口を跨ぐように、該開口の外周の異なる位置間を連結した把持部を設けている。そのため、細胞培養器具１０をウエル１１内に配置する際、あるいは、細胞培養器具１０をウエル１１から引き上げる際に、これらの把持部１２ｈまたは１４ｈを把持することによって細胞培養器具１０を吊り下げることが可能になり、細胞培養器具１０を取り扱う操作を容易化することができる。 Further, according to the cell culture instrument 10 of the present embodiment, the outer cylinder 12 or the first inner cylinder 14 is used as the first gripping part 12 h of the outer cylinder 12 and the second gripping part 14 h of the first inner cylinder 14. The grip part which connected between the different positions of the outer periphery of this opening is provided so that the opening of this edge part may be straddled. Therefore, when the cell culture instrument 10 is placed in the well 11 or when the cell culture instrument 10 is pulled up from the well 11, the cell culture instrument 10 can be suspended by gripping these gripping portions 12 h or 14 h. It becomes possible, and the operation which handles the cell culture instrument 10 can be made easy.

特に、本実施形態では、第１の把持部１２ｈおよび第２の把持部１４ｈを第１の内筒１４の軸線上で交わる２本の線分状に形成している。そのため、この２本の線分の交点で把持することにより、細胞培養器具１０の重心の直上付近において細胞培養器具１０を把持することが容易になる。その結果、細胞培養器具１０を吊り下げる際に、細胞培養用膜１６の水平を保ち、細胞培養用膜１６上で培養される細胞に対する望ましくない外的刺激（ノイズ）等を抑えることが容易になる。 In particular, in the present embodiment, the first grip portion 12h and the second grip portion 14h are formed in two line segments that intersect on the axis of the first inner cylinder 14. Therefore, by grasping at the intersection of the two line segments, it becomes easy to grasp the cell culture instrument 10 in the vicinity immediately above the center of gravity of the cell culture instrument 10. As a result, when the cell culture instrument 10 is suspended, it is easy to keep the cell culture membrane 16 horizontal and suppress unwanted external stimuli (noise), etc., on the cells cultured on the cell culture membrane 16. Become.

なお、第１の把持部１２ｈおよび第２の把持部１４ｈに係る上記２本の線分の交点が存在する「第１の内筒１４の軸線上」とは、第１の内筒１４の軸線上付近を含むものである。具体的には、「上記交点が第１の内筒１４の軸線上に存在する」とは、外筒１２および第１の内筒１４が円柱状である場合には、上記交点を含み第１の内筒１４の軸線に垂直な面において、上記交点が、第１の内筒１４の内周に比べて直径が１０分の１である第１の内筒１４の同心円の範囲内に含まれる構成を含む。また、後述するように、第１の内筒１４は円柱状以外の筒状に形成することが可能であるが、このような場合には、上記交点を含み第１の内筒１４の軸線に垂直な面において、上記交点が、第１の内筒１４の内周を重心が一致する状態で１０分の１に縮小させた範囲内に含まれる構成を含む。なお、細胞培養器具１０を吊り下げる際に、細胞培養膜１６の水平状態を保ち、細胞に対する望ましくない外的刺激（ノイズ）等を抑える観点からは、第１の内筒１４における軸線に垂直な面の形状は、回転対称であることが望ましい。 Note that “on the axis of the first inner cylinder 14” where the intersection of the two line segments related to the first gripping part 12h and the second gripping part 14h exists is the axis of the first inner cylinder 14 It includes the vicinity of the line. Specifically, “the intersection point exists on the axis of the first inner cylinder 14” means that when the outer cylinder 12 and the first inner cylinder 14 are cylindrical, the first point including the intersection point is included. In the plane perpendicular to the axis of the inner cylinder 14, the intersection point is included in the range of the concentric circle of the first inner cylinder 14 whose diameter is 1/10 of the inner circumference of the first inner cylinder 14. Includes configuration. Further, as will be described later, the first inner cylinder 14 can be formed in a cylinder shape other than the columnar shape. In such a case, the first inner cylinder 14 includes the intersection point and is on the axis of the first inner cylinder 14. In the vertical plane, the intersection includes a configuration in which the inner circumference of the first inner cylinder 14 is included in a range reduced to 1/10 in a state where the centers of gravity coincide with each other. Note that, when the cell culture instrument 10 is suspended, the cell culture membrane 16 is kept in a horizontal state, and from the viewpoint of suppressing undesirable external stimulation (noise) and the like to the cell, the cell culture instrument 10 is perpendicular to the axis of the first inner cylinder 14. The shape of the surface is preferably rotationally symmetric.

さらに、本実施形態の細胞培養器具１０によれば、外筒１２に形成される第１のスリット１２ｓは、第１の内筒１４の一方の端部１４ｔと外筒１２とが重なる位置から、第１の内筒１４から高さ方向に離間する方向に向かって延びている。したがって、外筒１２内の空間であって、細胞培養用膜１６の一方の面（最初に播種される細胞αの培養面）上の空間が、細胞αの培養用面の近傍において、第１のスリット１２ｓを介して外筒１２の外部と連通している。そのため、細胞培養器具１０を反転させた時に、細胞αの培養面から、第１のスリット１２ｓを介して、容易にエアを抜くことができる。図１（Ｂ）では、エアが抜かれる様子を矢印で示している。これにより、最初に播種される細胞αの培養面上にエアが残留することに起因する不都合（例えば、細胞αが培養液と接触しないこと）を抑えることができる。 Furthermore, according to the cell culture instrument 10 of the present embodiment, the first slit 12s formed in the outer cylinder 12 is located from the position where the one end portion 14t of the first inner cylinder 14 and the outer cylinder 12 overlap. It extends toward the direction away from the first inner cylinder 14 in the height direction. Therefore, the space in the outer cylinder 12 on one surface of the cell culture membrane 16 (the culture surface of the cell α to be seeded first) is the first in the vicinity of the cell α culture surface. The outer cylinder 12 communicates with the outside through the slit 12s. Therefore, when the cell culture instrument 10 is inverted, air can be easily extracted from the culture surface of the cell α via the first slit 12s. In FIG. 1B, the state in which air is extracted is indicated by arrows. Thereby, the inconvenience (for example, that cell alpha does not contact a culture solution) resulting from air remaining on the culture surface of cell alpha seeded initially can be controlled.

また、本実施形態の細胞培養器具１０によれば、細胞培養用膜１６として、厚み方向に貫通する多数の貫通孔を有する多孔質な膜を用いて、細胞培養用膜の両面において細胞を培養することができる。このような場合には、細胞培養器具１０を反転させるという簡便な操作により、細胞培養用膜１６の各々の面に細胞を播種する動作を容易に行なうことが可能になる。 Further, according to the cell culture instrument 10 of the present embodiment, the cell culture membrane 16 is a porous membrane having a large number of through holes penetrating in the thickness direction, and the cells are cultured on both surfaces of the cell culture membrane. can do. In such a case, the operation of seeding the cells on each surface of the cell culture membrane 16 can be easily performed by a simple operation of inverting the cell culture instrument 10.

Ｂ．第２の実施形態：
図４は、本発明の第２の実施形態である細胞培養器具１１０の構成を図１と同様にして表わす断面模式図であり、図５は、細胞培養器具１１０の組み立ての様子を説明するための分解斜視図である。以下の説明では、第１の実施形態の細胞培養器具１０と共通する部分には同じ参照番号を付して、詳しい説明は省略する。細胞培養器具１１０は、外筒１２と、第１の内筒１４と、細胞培養用膜１６と、に加えて、さらに第２の内筒１８を備える。 B. Second embodiment:
FIG. 4 is a schematic cross-sectional view showing the configuration of the cell culture instrument 110 according to the second embodiment of the present invention in the same manner as in FIG. 1, and FIG. 5 is a diagram for explaining how the cell culture instrument 110 is assembled. FIG. In the following description, parts common to the cell culture instrument 10 of the first embodiment are denoted by the same reference numerals, and detailed description thereof is omitted. The cell culture instrument 110 further includes a second inner cylinder 18 in addition to the outer cylinder 12, the first inner cylinder 14, and the cell culture membrane 16.

第２の内筒１８は、第１の内筒１４と同様の外径を有する筒状の部材であって、外筒１２よりも高さが小さく形成されている。第２の内筒１８は、第１の内筒１４と同様の材料により形成することができる。図４に示すように、第２の内筒１８は、第１の内筒１４と高さ方向が同じであって、第１の内筒１４における一方の端部１４ｏ側で細胞培養用膜１６を間に介して第１の内筒１４に接するように、外筒１２内に配置される。なお、第１の内筒１４と外筒１２とが細胞培養用膜１６を挟持する際の把持力を損なわないように、第２の内筒１８の外径は、第１の内筒１４の外径を超えないように形成されている。 The second inner cylinder 18 is a cylindrical member having an outer diameter similar to that of the first inner cylinder 14, and is formed with a height smaller than that of the outer cylinder 12. The second inner cylinder 18 can be formed of the same material as the first inner cylinder 14. As shown in FIG. 4, the second inner cylinder 18 has the same height direction as the first inner cylinder 14, and the cell culture membrane 16 on the one end portion 14 o side of the first inner cylinder 14. It arrange | positions in the outer cylinder 12 so that the 1st inner cylinder 14 may be contacted via. The outer diameter of the second inner cylinder 18 is set so that the gripping force when the first inner cylinder 14 and the outer cylinder 12 sandwich the cell culture membrane 16 is not impaired. It is formed so as not to exceed the outer diameter.

図５に示すように、細胞培養器具１１０を組み立てる際には、第１の内筒１４の一方の端部１４ｏ（第２の把持部１４ｈを有しない側の端部）上に細胞培養用膜１６を配置する際に、細胞培養用膜１６上にさらに第２の内筒１８を配置する点が、第１の実施形態と異なっている。その後、第１の内筒１４に対する嵌め込みの方向が第１の実施形態と同様になるように、細胞培養用膜１６を挟持する第１の内筒１４および第２の内筒１８の外側に、外筒１２を嵌め込む。 As shown in FIG. 5, when the cell culture instrument 110 is assembled, the cell culture membrane is placed on one end portion 14o of the first inner cylinder 14 (the end portion on the side not having the second gripping portion 14h). The point which arrange | positions the 2nd inner cylinder 18 on the film | membrane 16 for cell cultures when arrange | positioning 16 differs from 1st Embodiment. Thereafter, outside the first inner cylinder 14 and the second inner cylinder 18 sandwiching the cell culture membrane 16 so that the fitting direction with respect to the first inner cylinder 14 is the same as in the first embodiment, The outer cylinder 12 is fitted.

図４（Ａ）は、図１（Ａ）と同様に、細胞培養用膜１６の一方の面上に細胞αを播種した様子を表わす。また、図４（Ｂ）は、図４（Ａ）の細胞培養器具１１０を反転させて、図１（Ｂ）と同様に細胞培養用膜１６の他方の面上に、細胞αとは異なる細胞βをさらに播種した様子を示す。本実施形態では、内部に第１の内筒１４を収容したときの外筒１２の内径と第２の内筒１８の外径の関係、外筒１２の内壁面と第２の内筒１８の外側面の表面粗さ、および第２の内筒１８の重量とを調節することにより、反転後であっても外筒１２に対する第２の内筒１８の相対的な位置を維持可能となっている。 FIG. 4A shows a state in which cells α are seeded on one surface of the cell culture membrane 16 as in FIG. 1A. 4B shows a cell different from the cell α on the other surface of the cell culture membrane 16 as in FIG. 1B by inverting the cell culture instrument 110 of FIG. 4A. A state where β is further seeded is shown. In the present embodiment, the relationship between the inner diameter of the outer cylinder 12 and the outer diameter of the second inner cylinder 18 when the first inner cylinder 14 is accommodated therein, the inner wall surface of the outer cylinder 12 and the second inner cylinder 18. By adjusting the surface roughness of the outer surface and the weight of the second inner cylinder 18, the relative position of the second inner cylinder 18 with respect to the outer cylinder 12 can be maintained even after inversion. Yes.

このような構成としても、第１の実施形態と同様に、外筒１２に第１のスリット１２ｓを設けたことで細胞培養用膜１６を容易に保持できるという既述した効果が得られる。さらに、第２の実施形態では、第２の内筒１８をさらに用いるため、細胞αを播種する際に、細胞培養用膜１６上に供給した細胞αが、細胞培養器具１１０の外部に流出することを抑えられる。そのため、細胞αを播種する際に、播種する細胞数の精度を向上させることができる。 Even in such a configuration, as described in the first embodiment, by providing the first slit 12s in the outer cylinder 12, the above-described effect that the cell culture membrane 16 can be easily held is obtained. Furthermore, in the second embodiment, since the second inner cylinder 18 is further used, when the cells α are seeded, the cells α supplied onto the cell culture membrane 16 flow out of the cell culture instrument 110. It can be suppressed. Therefore, the accuracy of the number of cells to be seeded can be improved when the cells α are seeded.

Ｃ．第３の実施形態：
図６は、本発明の第３の実施形態である細胞培養器具２１０の構成を図１と同様にして表わす断面模式図である。以下の説明では、第１の実施形態の細胞培養器具１０と共通する部分には同じ参照番号を付して、詳しい説明は省略する。細胞培養器具２１０は、第１の内筒１４に代えて第１の内筒２１４を備える以外は、細胞培養器具１０と同様の構成を有している。第１の内筒２１４には、第２のスリット２１４ｓおよび一対の膜支持部２１４ｍが設けられている。 C. Third embodiment:
FIG. 6 is a schematic cross-sectional view showing the configuration of a cell culture instrument 210 according to the third embodiment of the present invention in the same manner as FIG. In the following description, parts common to the cell culture instrument 10 of the first embodiment are denoted by the same reference numerals, and detailed description thereof is omitted. The cell culture instrument 210 has the same configuration as that of the cell culture instrument 10 except that the first inner cylinder 214 is provided instead of the first inner cylinder 14. The first inner cylinder 214 is provided with a second slit 214s and a pair of membrane support portions 214m.

図７は、第１の内筒２１４の一部を拡大して示す斜視図である。第２のスリット２１４ｓは、第１の内筒２１４において、一方の端部１４ｏ（第２の把持部１４ｈを有しない側であって細胞培養用膜１６が配置される側の端部。図６参照）から高さ方向に延びるように形成されている。一対の膜支持部２１４ｍは、第１の内筒２１４の一方の端部１４ｏにおける第２のスリット２１４ｓが開口する位置において、第１の内筒２１４の径方向外側に向かって延びるように設けられている。第１の内筒２１４は、第２のスリット２１４ｓおよび膜支持部２１４ｍが、外筒１２に設けた第１のスリット１２ｓと重なり、膜支持部２１４ｍが、第１のスリット１２ｓを経由して外筒１２の外側に延びるように、外筒１２内に配置される。 FIG. 7 is an enlarged perspective view showing a part of the first inner cylinder 214. In the first inner cylinder 214, the second slit 214s is one end portion 14o (the end portion on the side not having the second gripping portion 14h on which the cell culture membrane 16 is disposed). It is formed to extend in the height direction from the reference). The pair of membrane support portions 214m are provided so as to extend outward in the radial direction of the first inner cylinder 214 at a position where the second slit 214s is opened at one end portion 14o of the first inner cylinder 214. ing. In the first inner cylinder 214, the second slit 214s and the membrane support portion 214m overlap with the first slit 12s provided in the outer cylinder 12, and the membrane support portion 214m is outside via the first slit 12s. It arrange | positions in the outer cylinder 12 so that the outer side of the cylinder 12 may be extended.

細胞培養器具２１０を組み立てる際には、第１の実施形態と同様に、第１の内筒２１４の一方の端部１４ｏ上に細胞培養用膜１６を配置して、この一方の端部１４ｏ側から、外筒１２の他方の端部１２ｔ（第１の把持部１２ｈを有しない側の端部）を嵌め込む。これにより、細胞培養用膜１６を、第１の内筒２１４の一方の端部１４ｏの外周に沿って折り曲げて、細胞培養用膜１６の周縁部を、第１の内筒２１４の外側面と外筒１２の内壁面によって挟持する。このとき、第１の内筒２１４には膜支持部２１４ｍが設けられているため、膜支持部２１４ｍの近傍では、細胞培養用膜１６が第１の内筒２１４の外側面に接することがない。その結果、第１の内筒２１４の第２のスリット２１４ｓが細胞培養用膜１６に塞がれることなく、第１の内筒２１４の内部と外筒１２の外部とは、第１のスリット１２ｓおよび第２のスリット２１４ｓを介して連通する。 When assembling the cell culture instrument 210, as in the first embodiment, the cell culture membrane 16 is arranged on one end portion 14o of the first inner cylinder 214, and this one end portion 14o side is arranged. Then, the other end portion 12t of the outer cylinder 12 (the end portion on the side not having the first gripping portion 12h) is fitted. Thereby, the cell culture membrane 16 is bent along the outer periphery of the one end portion 14o of the first inner cylinder 214, and the peripheral portion of the cell culture membrane 16 is connected to the outer surface of the first inner cylinder 214. It is clamped by the inner wall surface of the outer cylinder 12. At this time, since the first inner cylinder 214 is provided with the membrane support portion 214m, the cell culture membrane 16 does not contact the outer surface of the first inner cylinder 214 in the vicinity of the membrane support portion 214m. . As a result, the second slit 214s of the first inner cylinder 214 is not blocked by the cell culture membrane 16, and the inside of the first inner cylinder 214 and the outside of the outer cylinder 12 are separated from each other by the first slit 12s. And it communicates via the second slit 214s.

このような構成としても、第１の実施形態と同様に、外筒１２に第１のスリット１２ｓを設けたことで細胞培養用膜１６を容易に保持できるという既述した効果が得られる。さらに、第３の実施形態では、第１の内筒２１４に第２のスリット２１４ｓおよび膜支持部２１４ｍを設けて第１の内筒２１４内と外筒１２の外部とを連通させている。そのため、図６の状態で細胞培養を行なう際に、細胞培養用膜１６の下側に残留するエアを、容易に排出することができる。図６では、エアが抜かれる様子を矢印で示している。これにより、図１（Ａ）と同様に細胞αを播種して培養する際に、培養面の裏面にエアが残留することに起因する不都合を抑えることができる。さらに本実施形態では、図６の状態から細胞培養器具２１０を上下反転させて、図１（Ｂ）と同様に細胞βを培養する際にも、図１（Ｂ）と同様に、細胞αの培養面から第１のスリット１２ｓを介して容易にエアを抜くことができる。 Even in such a configuration, as described in the first embodiment, by providing the first slit 12s in the outer cylinder 12, the above-described effect that the cell culture membrane 16 can be easily held is obtained. Furthermore, in the third embodiment, the first inner cylinder 214 is provided with a second slit 214s and a film support portion 214m so that the inside of the first inner cylinder 214 communicates with the outside of the outer cylinder 12. Therefore, when the cell culture is performed in the state of FIG. 6, the air remaining below the cell culture membrane 16 can be easily discharged. In FIG. 6, the state where air is extracted is indicated by arrows. Thereby, when seed | inoculating and culture | cultivating the cell (alpha) like FIG. 1 (A), the inconvenience resulting from air remaining on the back surface of a culture surface can be suppressed. Further, in this embodiment, when the cell culture instrument 210 is turned upside down from the state of FIG. 6 and the cell β is cultured in the same manner as in FIG. Air can be easily extracted from the culture surface via the first slit 12s.

なお、第３の実施形態では、第２のスリット２１４ｓは、第１の内筒２１４の一方の端部１４ｏから、第１の内筒２１４の高さ方向の途中の位置まで延びる切れ込み状の形状としたが、異なる形状としてもよい。例えば、第１の内筒２１４の高さ方向全体にわたって延びるように、第２のスリット２１４ｓを設けてもよい。第２のスリット２１４ｓは、細胞αの培養面の裏面の直上付近の空間が、外部と連通可能となる形状であればよい。 In the third embodiment, the second slit 214 s has a notch shape extending from one end portion 14 o of the first inner cylinder 214 to a position in the middle of the first inner cylinder 214 in the height direction. However, it may have a different shape. For example, the second slit 214s may be provided so as to extend over the entire height direction of the first inner cylinder 214. The second slit 214s only needs to have a shape in which the space immediately above the rear surface of the culture surface of the cell α can communicate with the outside.

また、第３の実施形態では、図７に示すように、第１の内筒２１４には、一方の端部１４ｏ近傍の外側面であって、細胞培養用膜１６の周縁部と接する位置に、第１の内筒２１４の側面の周方向に沿って延びる細長い凹部である２本の溝２１４ｃが設けられている。このような構成とすることで、細胞培養用膜１６の周縁部を第１の内筒２１４および外筒１２によって挟持する際の保持力を高めることができる。なお、第１の内筒２１４の外側面に設ける構造は、上記２本の溝状構造のような凹部のみによって形成する他、凸部のみにより形成してもよく、凹部と凸部の組合せにより形成してもよく、細胞培養用膜１６の保持力を高めることができる凹凸形状であればよい。また、このような凹凸形状は、他のいずれの実施形態や変形例に適用してもよい。 In the third embodiment, as shown in FIG. 7, the first inner cylinder 214 has an outer surface in the vicinity of one end portion 14 o at a position in contact with the peripheral portion of the cell culture membrane 16. Two grooves 214c, which are elongated concave portions extending along the circumferential direction of the side surface of the first inner cylinder 214, are provided. With such a configuration, it is possible to increase the holding force when the peripheral portion of the cell culture membrane 16 is sandwiched between the first inner tube 214 and the outer tube 12. The structure provided on the outer surface of the first inner cylinder 214 may be formed only by a concave portion such as the above two groove-like structures, or may be formed only by a convex portion, or by a combination of the concave and convex portions. It may be formed as long as it has a concavo-convex shape that can enhance the retention of the cell culture membrane 16. Moreover, you may apply such uneven | corrugated shape to any other embodiment and modification.

Ｄ．第４の実施形態：
図８は、本発明の第４の実施形態である細胞培養器具３１０の構成を、図１と同様にして表わす断面模式図である。以下の説明では、第１〜第３の実施形態と共通する部分には同じ参照番号を付して、詳しい説明は省略する。細胞培養器具３１０は、外筒１２と、第１の内筒２１４と、細胞培養用膜１６と、第２の内筒３１８とを備える。 D. Fourth embodiment:
FIG. 8 is a schematic cross-sectional view showing the configuration of a cell culture instrument 310 according to the fourth embodiment of the present invention in the same manner as FIG. In the following description, parts common to the first to third embodiments are denoted by the same reference numerals, and detailed description thereof is omitted. The cell culture instrument 310 includes an outer cylinder 12, a first inner cylinder 214, a cell culture membrane 16, and a second inner cylinder 318.

第２の内筒３１８は、高さ方向が同じになるように第１の内筒２１４と重ねて、第１の内筒２１４と共に細胞培養用膜１６を端面で挟持するように配置される。この第２の内筒３１８は、細胞培養用膜１６を対称面として、第１の内筒２１４と面対称となる形状を有している。すなわち、第２の内筒３１８には、第２のスリット２１４ｓと同様の第３のスリット３１８ｓ、および、膜支持部２１４ｍと同様の膜支持部３１８ｍが設けられている。 The second inner cylinder 318 is placed so as to overlap the first inner cylinder 214 so that the height direction is the same, and sandwich the cell culture membrane 16 with the first inner cylinder 214 at the end face. The second inner cylinder 318 has a shape that is symmetrical with the first inner cylinder 214 with the cell culture membrane 16 as a symmetry plane. That is, the second inner cylinder 318 is provided with a third slit 318s similar to the second slit 214s and a membrane support 318m similar to the membrane support 214m.

このような構成とすれば、第３の実施形態の細胞培養器具２１０と同様の効果を奏すると共に、さらに、細胞αを播種して培養した後、細胞培養器具３１０を上下反転させたときに、細胞αの培養面の直上に残留するエアを、第３のスリット３１８ｓを介して容易に排出可能となる。なお、第３のスリット３１８ｓの形状は、第２のスリット２１４ｓと同じ形状である必要はなく、細胞αの培養面の直上付近の空間が外部と連通可能となる形状であればよい。 With such a configuration, the same effect as the cell culture instrument 210 of the third embodiment is obtained, and further, when the cell culture instrument 310 is turned upside down after seeding and culturing the cells α, The air remaining immediately above the culture surface of the cell α can be easily discharged through the third slit 318s. Note that the shape of the third slit 318s is not necessarily the same shape as the second slit 214s, and may be any shape as long as the space immediately above the culture surface of the cell α can communicate with the outside.

Ｅ．第５の実施形態：
図９は、本発明の第５の実施形態である細胞培養器具４１０の構成を表わす断面模式図である。以下の説明では、第１の実施形態の細胞培養器具１０と共通する部分には同じ参照番号を付して、詳しい説明は省略する。細胞培養器具４１０は、外筒１２、第１の内筒１４、および細胞培養用膜１６に加えて、さらに第２の内筒４１８を備えている。 E. Fifth embodiment:
FIG. 9 is a schematic cross-sectional view showing the configuration of a cell culture instrument 410 according to the fifth embodiment of the present invention. In the following description, parts common to the cell culture instrument 10 of the first embodiment are denoted by the same reference numerals, and detailed description thereof is omitted. The cell culture instrument 410 is further provided with a second inner cylinder 418 in addition to the outer cylinder 12, the first inner cylinder 14, and the cell culture membrane 16.

第２の内筒４１８は、高さ方向が同じになるように第１の内筒１４と重ねて、第１の内筒１４と共に細胞培養用膜１６を端面で挟持するように配置される。なお、本実施形態では、外筒１２には第１の把持部１２ｈが設けられておらず、細胞培養用膜１６から離間する側の第２の内筒４１８の端部において、第１の把持部１２ｈと同様の第３の把持部４１８ｈが設けられている。 The second inner cylinder 418 is disposed so as to overlap the first inner cylinder 14 so that the height direction is the same, and sandwich the cell culture membrane 16 with the first inner cylinder 14 at the end face. In the present embodiment, the outer cylinder 12 is not provided with the first gripping portion 12h, and the first gripping is performed at the end of the second inner tube 418 on the side away from the cell culture membrane 16. A third grip portion 418h similar to the portion 12h is provided.

図９（Ａ）は、図１（Ａ）と同様に、細胞培養用膜１６の一方の面（第２の内筒４１８の内部に露出する面）上に細胞αを播種した様子を表わす。図９（Ｂ）は、図９（Ａ）の状態から、第２の内筒４１８が細胞培養用膜１６から離間するように高さ方向（鉛直方向上方）に第２の内筒４１８を引き上げた様子を表わす。このような第２の内筒４１８の引き上げは、第３の把持部４１８ｈを把持することで、容易に行なうことができる。図９（Ｃ）は、図９（Ｂ）の状態から細胞培養器具４１０を上下反転させて、細胞αの培養面の裏面に、さらに細胞βを播種した様子を表わす。細胞培養器具４１０を上下反転させる動作も、第３の把持部４１８ｈを把持して細胞培養器具４１０を吊り下げつつ引き上げることにより、容易に行なうことができる。 FIG. 9A shows a state in which cells α are seeded on one surface of the cell culture membrane 16 (the surface exposed inside the second inner cylinder 418), as in FIG. 1A. 9B, the second inner cylinder 418 is pulled up in the height direction (vertically upward) so that the second inner cylinder 418 is separated from the cell culture membrane 16 from the state of FIG. 9A. Represents the appearance. Such pulling up of the second inner cylinder 418 can be easily performed by holding the third holding portion 418h. FIG. 9C shows a state in which the cell culture instrument 410 is turned upside down from the state of FIG. 9B and the cells β are further seeded on the back surface of the culture surface of the cells α. The operation of turning the cell culture instrument 410 upside down can also be easily performed by grasping the third gripping part 418h and pulling up the cell culture instrument 410 while hanging.

本実施形態では、第２の内筒４１８は、外筒１２を押さえつつ第２の内筒４１８に対して高さ方向上方の力を加えることにより、外筒１２内で第２の内筒４１８を高さ方向にスライドさせて、スライドさせた位置にて保持可能となっている。このような構成は、内部に第１の内筒１４を収容したときの外筒１２の内径と第２の内筒４１８の外径の関係、外筒１２の内壁面と第２の内筒４１８の外側面の表面粗さ、および第２の内筒４１８の重量とを調節することにより、実現可能である。 In the present embodiment, the second inner cylinder 418 applies a force in the height direction to the second inner cylinder 418 while pressing the outer cylinder 12, thereby causing the second inner cylinder 418 within the outer cylinder 12. Is slid in the height direction and can be held at the slid position. In such a configuration, the relationship between the inner diameter of the outer cylinder 12 and the outer diameter of the second inner cylinder 418 when the first inner cylinder 14 is accommodated therein, the inner wall surface of the outer cylinder 12 and the second inner cylinder 418. This can be realized by adjusting the surface roughness of the outer surface and the weight of the second inner cylinder 418.

このような構成としても、第１の実施形態と同様に、外筒１２に第１のスリット１２ｓを設けたことで細胞培養用膜１６を容易に保持できるという既述した効果が得られる。さらに、第５の実施形態では、細胞培養器具４１０の反転に先立って第２の内筒４１８を高さ方向にスライドさせて細胞培養用膜１６から離間させることで、細胞αの培養面上の直上の空間が、外筒１２の第１のスリット１２ｓを介して外筒１２の外部と連通する。そのため、細胞培養器具４１０の反転後には、細胞αの培養面上に残留するエアを、第１のスリット１２ｓを介して容易に排出することができる。図９（Ｃ）では、エアが抜かれる様子を矢印で示している。これにより、細胞αの培養面にエアが残留することに起因する不都合を抑えることができる。 Even in such a configuration, as described in the first embodiment, by providing the first slit 12s in the outer cylinder 12, the above-described effect that the cell culture membrane 16 can be easily held is obtained. Furthermore, in the fifth embodiment, the second inner cylinder 418 is slid in the height direction and separated from the cell culture membrane 16 prior to the reversal of the cell culture instrument 410, so that the cell α is on the culture surface. The space immediately above communicates with the outside of the outer cylinder 12 via the first slit 12 s of the outer cylinder 12. Therefore, after the cell culture instrument 410 is inverted, the air remaining on the culture surface of the cell α can be easily discharged through the first slit 12s. In FIG. 9C, the state in which air is extracted is indicated by arrows. Thereby, the inconvenience resulting from air remaining on the culture surface of the cell α can be suppressed.

なお、本実施形態では、第１の内筒１４と第２の内筒４１８の高さの合計は、外筒１２の高さよりも短いが、異なる構成としてもよい。細胞培養器具４１０を反転させた後にも、第２の内筒４１８をスライドさせた位置にて保持可能であればよい。また、外筒１２内での第２の内筒４１８の保持は、両者の間の摩擦力を利用するのではなく、例えば、外筒１２の内壁面と第２の内筒４１８の外側面に、両者が係合可能となる係合部を設けてもよい。ただし、繰り返し任意の位置に第２の内筒４１８をスライド可能であるという観点からは、上記摩擦力を利用することが望ましい。 In the present embodiment, the total height of the first inner cylinder 14 and the second inner cylinder 418 is shorter than the height of the outer cylinder 12, but may have a different configuration. Even if the cell culture instrument 410 is inverted, it is sufficient if the second inner cylinder 418 can be held at the slid position. Further, the holding of the second inner cylinder 418 in the outer cylinder 12 does not use the frictional force between them, but, for example, on the inner wall surface of the outer cylinder 12 and the outer surface of the second inner cylinder 418. In addition, an engaging portion that enables both to be engaged may be provided. However, from the viewpoint that the second inner cylinder 418 can be slid repeatedly at an arbitrary position, it is desirable to use the friction force.

Ｆ．変形例：
・変形例１：
上記各実施形態では、外筒１２に設けた第１のスリット１２ｓは、外筒１２の高さ方向全体にわたって形成されている。これに対して、第１のスリット１２ｓは、外筒１２の高さ方向の一部の領域に形成することとしてもよい。第１のスリット１２ｓは、外筒１２の高さ方向に沿って延びており、少なくとも、細胞培養用膜１６が配置される第１の内筒の端部と重なる位置に設けられていれば、第１の内筒と共に細胞培養用膜１６を挟持することにより、細胞培養用膜１６を保持するための把持力を生じることができる。 F. Variations:
・ Modification 1:
In each of the above embodiments, the first slit 12 s provided in the outer cylinder 12 is formed over the entire height direction of the outer cylinder 12. On the other hand, the first slit 12s may be formed in a partial region of the outer cylinder 12 in the height direction. If the first slit 12s extends along the height direction of the outer cylinder 12 and is provided at least at a position overlapping the end of the first inner cylinder where the cell culture membrane 16 is disposed, By holding the cell culture membrane 16 together with the first inner cylinder, a gripping force for holding the cell culture membrane 16 can be generated.

なお、外筒１２において高さ方向の一部の領域のみに第１のスリット１２ｓを設ける場合には、外筒１２に対する第１の内筒１４等の挿入を容易にするために、例えば、内筒の挿入側先端部に、先端に向かって縮径するテーパを設けることが好ましい。あるいは、外筒１２において、第１のスリット１２ｓが設けられる領域以外の領域では、第１のスリット１２ｓが設けられる領域に比べて外筒１２の外径を若干大きくして、外筒１２への第１の内筒１４等の嵌め込みを容易化してもよい。 In the case where the first slit 12s is provided only in a partial region in the height direction of the outer cylinder 12, in order to facilitate the insertion of the first inner cylinder 14 and the like into the outer cylinder 12, for example, It is preferable to provide a taper that decreases in diameter toward the tip at the tip on the insertion side of the tube. Alternatively, in the outer cylinder 12, in the area other than the area where the first slit 12s is provided, the outer diameter of the outer cylinder 12 is slightly larger than the area where the first slit 12s is provided, The first inner cylinder 14 and the like may be easily fitted.

・変形例２：
上記実施形態では、外筒１２、第１の内筒、および第２の内筒に設けた把持部は、いずれも上面視を十字形状としたが、異なる形状としてもよい。把持部の形状は、例えば、外筒あるいは内筒の端部の開口を跨ぐように、該開口の外周の異なる位置間を連結した形状のように、細胞培養器具の把持の容易化が可能であればよい。細胞培養器具の重心の直上付近（軸線上付近）で器具を把持可能となる形状であれば、細胞培養器具を安定して保持し易く、培養細胞に対する外的刺激（ノイズ）を抑え易くなる観点から、特に望ましい。ただし、各実施形態で説明した把持部のうちの少なくとも一部を設けない構成も可能である。 Modification 2
In the above embodiment, the gripping portions provided on the outer cylinder 12, the first inner cylinder, and the second inner cylinder are all cross-shaped in top view, but may have different shapes. The shape of the gripping part can facilitate gripping of the cell culture instrument, for example, a shape in which different positions on the outer periphery of the opening are connected so as to straddle the opening of the end of the outer cylinder or the inner cylinder. I just need it. If the shape allows the device to be gripped near the center of gravity of the cell culture device (near the axis), it is easy to stably hold the cell culture device and to suppress external stimulation (noise) to the cultured cells. Particularly desirable. However, the structure which does not provide at least one part of the holding part demonstrated by each embodiment is also possible.

なお、培養細胞に対する外的刺激の低減の観点からは、細胞培養器具は培養液中に沈むことが望ましい。細胞培養器具の比重が十分に大きくない場合には、培養液中に沈んだ状態で細胞培養器具を保持する機構を設けることが望ましい。 Note that, from the viewpoint of reducing external stimulation to the cultured cells, it is desirable that the cell culture device is submerged in the culture solution. When the specific gravity of the cell culture device is not sufficiently large, it is desirable to provide a mechanism for holding the cell culture device in a state of being submerged in the culture solution.

・変形例３：
上記実施形態では、細胞培養用膜の両面を用いて細胞培養することとしたが、片面のみを用いて細胞培養してもよい。この場合には、細胞培養用膜の両面間が貫通孔によって連通している必要はないため、例えばＰＥＴ等の樹脂の薄膜から成る基板上に、培養細胞の足場となる層を成膜してもよい。すなわち、培養細胞の足場となる層自身が、十分な弾性および強度を有していなくても、基板を含めた全体で、第１の内筒の端部で折れ曲げられて、第１の内筒と外筒の間で挟持されるための十分な弾性および強度を有していればよい。・ Modification 3:
In the above embodiment, cell culture is performed using both surfaces of the cell culture membrane, but cell culture may be performed using only one surface. In this case, both sides of the cell culture membrane do not need to communicate with each other by a through-hole. For example, a layer to be a scaffold for cultured cells is formed on a substrate made of a resin thin film such as PET. Also good. That is, even if the layer itself serving as a scaffold for the cultured cells does not have sufficient elasticity and strength, it is bent at the end of the first inner cylinder as a whole including the substrate, and the first inner cylinder is bent. It is only necessary to have sufficient elasticity and strength to be sandwiched between the cylinder and the outer cylinder.

細胞培養器具を片面培養に用いる場合には、細胞培養用膜の裏面に対して、播種のために細胞を懸濁した培養液を供給する必要がない。そのため、上記各実施形態のように、外筒１２の高さ方向の中ほどに細胞培養用膜１６を配置するのではなく、外筒１２の他方の端部１４ｔの近傍に細胞培養用膜１６を配置してもよい。 When the cell culture instrument is used for single-sided culture, it is not necessary to supply a culture solution in which cells are suspended for seeding to the back surface of the cell culture membrane. Therefore, the cell culture membrane 16 is not arranged in the middle of the height direction of the outer cylinder 12 as in the above embodiments, but the cell culture membrane 16 is provided in the vicinity of the other end portion 14t of the outer cylinder 12. May be arranged.

・変形例４：
上記各実施形態では、外筒および内筒（第１の内筒、第２の内筒）を円筒状としたが、異なる構成としてもよい。例えば、内筒を円筒状に形成し、外筒を、この内筒に外接する多角柱状（例えば六角柱や八角柱）としてもよい。また、外筒を円柱状に形成し、内筒を、この外筒に内接する多角柱状としてもよい。あるいは、内筒と外筒の双方を、多角柱状とすることも可能である。内筒の外側面と外筒の内壁面とは、周方向全体にわたって接する必要はなく、細胞培養膜を保持するために十分な把持力が生じる程度に接していればよい。 -Modification 4:
In each of the above embodiments, the outer cylinder and the inner cylinder (the first inner cylinder and the second inner cylinder) are cylindrical, but may have different configurations. For example, the inner cylinder may be formed in a cylindrical shape, and the outer cylinder may have a polygonal column shape (for example, a hexagonal column or an octagonal column) circumscribing the inner cylinder. Further, the outer cylinder may be formed in a columnar shape, and the inner cylinder may be formed in a polygonal column shape inscribed in the outer cylinder. Or it is also possible to make both an inner cylinder and an outer cylinder into a polygonal column shape. The outer surface of the inner cylinder and the inner wall surface of the outer cylinder need not be in contact with each other in the entire circumferential direction, as long as sufficient gripping force is generated to hold the cell culture membrane.

本発明は、上述の実施形態や実施例、変形例に限られるものではなく、その趣旨を逸脱しない範囲において種々の構成で実現することができる。例えば、発明の概要の欄に記載した各形態中の技術的特徴に対応する実施形態、実施例、変形例中の技術的特徴は、上述の課題の一部又は全部を解決するために、あるいは、上述の効果の一部又は全部を達成するために、適宜、差し替えや、組み合わせを行うことが可能である。また、その技術的特徴が本明細書中に必須なものとして説明されていなければ、適宜、削除することが可能である。 The present invention is not limited to the above-described embodiments, examples, and modifications, and can be realized with various configurations without departing from the spirit thereof. For example, the technical features in the embodiments, examples, and modifications corresponding to the technical features in each embodiment described in the summary section of the invention are to solve some or all of the above-described problems, or In order to achieve part or all of the above effects, replacement or combination can be performed as appropriate. Further, if the technical feature is not described as essential in the present specification, it can be deleted as appropriate.

１０，１１０，２１０，３１０，４１０…細胞培養器具
１１…ウエル
１２…外筒
１２ｈ…第１の把持部
１２ｏ，１２ｔ…端部
１２ｓ…第１のスリット
１４，２１４…第１の内筒
１４ｈ…第２の把持部
１４ｏ，１４ｔ…端部
１６…細胞培養用膜
１８，３１８，４１８…第２の内筒
２１４ｃ…溝
２１４ｍ，３１８ｍ…膜支持部
２１４ｓ…第２のスリット
３１８ｓ…第３のスリット
４１８ｈ…第３の把持部 10, 110, 210, 310, 410 ... cell culture device 11 ... well 12 ... outer cylinder 12h ... first gripping part 12o, 12t ... end 12s ... first slit 14, 214 ... first inner cylinder 14h ... Second gripping part 14o, 14t ... End 16 ... Cell culture membrane 18, 318, 418 ... Second inner cylinder 214c ... Groove 214m, 318m ... Membrane support 214s ... Second slit 318s ... Third slit 418h ... third gripping portion

本発明は、上述の課題の少なくとも一部を解決するためになされたものであり、以下の形態として実現することが可能である。
本発明の一形態は、細胞培養器具であって；筒状の第１の内筒であって、該第１の内筒の一方の端部全体を覆うように細胞培養用膜を配置するための第１の内筒と；筒状に形成されて、前記第１の内筒と高さ方向が一致するように前記第１の内筒を内部に収容する外筒であって、少なくとも前記第１の内筒の前記一方の端部と重なる領域において、前記高さ方向に延び、前記外筒を径方向に弾性変形可能にする第１のスリットが形成されると共に、前記第１の内筒が内部にはめ込まれたときには、前記外筒の弾性復元力により、前記第１の内筒を保持する外筒と；を備える。このような形態であれば、第１の内筒の一方の端部と重なる領域において高さ方向に延びる第１のスリットを設けて、外筒の弾性復元力により、一方の端部全体を覆うように細胞培養用膜を配置するための第１の内筒を保持するため、各部材の寸法精度に対する要求を抑えつつ十分に高い信頼性で、細胞培養用膜を保持することが可能になる。その結果、細胞培養器具の製造工程の煩雑化やコストの増加を抑えることができる。また、第１の内筒の一方の端部と重なる領域において高さ方向に延びる第１のスリットを設けるため、第１の内筒の一方の端部全体を覆うように細胞培養用膜を配置し、細胞培養用膜上に細胞を播種した後に、細胞培養器具を上下反転させたときに、細胞培養用膜における上記細胞を播種した面上に残留するエアを、第１のスリットを介して排出させることが可能になる。
その他、本発明は、以下のような形態として実現することも可能である。 SUMMARY An advantage of some aspects of the invention is to solve at least a part of the problems described above, and the invention can be implemented as the following forms.
One aspect of the present invention is a cell culture instrument; a cylindrical first inner cylinder for disposing a cell culture membrane so as to cover the entire one end of the first inner cylinder. An outer cylinder that is formed in a cylindrical shape and accommodates the first inner cylinder so that the height direction of the first inner cylinder coincides with at least the first inner cylinder. A first slit extending in the height direction and elastically deforming the outer cylinder in a radial direction in a region overlapping with the one end of the one inner cylinder; and the first inner cylinder And an outer cylinder that holds the first inner cylinder by an elastic restoring force of the outer cylinder. If it is such a form, the 1st slit extended in a height direction in the area | region which overlaps with the one edge part of a 1st inner cylinder will be provided, and one whole edge part will be covered with the elastic restoring force of an outer cylinder As described above, since the first inner cylinder for arranging the cell culture membrane is held, it becomes possible to hold the cell culture membrane with sufficiently high reliability while suppressing the demand for dimensional accuracy of each member. . As a result, the complexity of the manufacturing process of the cell culture instrument and the increase in cost can be suppressed. Further, in order to provide a first slit extending in the height direction in a region overlapping with one end portion of the first inner cylinder, a cell culture membrane is disposed so as to cover the entire one end portion of the first inner cylinder. After the cells are seeded on the cell culture membrane, when the cell culture instrument is turned upside down, the air remaining on the surface of the cell culture membrane seeded with the cells is passed through the first slit. It becomes possible to discharge.
In addition, the present invention can be realized in the following forms.

Claims

細胞培養器具であって、
筒状の第１の内筒と、
前記第１の内筒の一方の端部全体を覆って配置された細胞培養用膜であって、前記第１の内筒の前記一方の端部の外周に沿って、前記第１の内筒の外側面側へと折り曲げられた細胞培養用膜と、
筒状に形成されて、前記第１の内筒と高さ方向が一致するように前記第１の内筒を内部に収容する外筒であって、少なくとも前記第１の内筒の前記一方の端部と重なる領域において、前記高さ方向に延びる第１のスリットが形成されると共に、前記外筒の内壁面と前記第１の内筒の外側面との間で、前記細胞培養用膜を挟持する外筒と、
を備える細胞培養器具。 A cell culture instrument,
A cylindrical first inner cylinder;
A membrane for cell culture arranged so as to cover the entire one end of the first inner cylinder, wherein the first inner cylinder extends along the outer periphery of the one end of the first inner cylinder. A cell culture membrane folded to the outer surface side of
An outer cylinder that is formed in a cylindrical shape and accommodates the first inner cylinder so that the height direction of the first inner cylinder coincides with at least one of the first inner cylinders. A first slit extending in the height direction is formed in a region overlapping with the end, and the cell culture membrane is disposed between the inner wall surface of the outer tube and the outer surface of the first inner tube. An outer cylinder to sandwich,
A cell culture instrument comprising:

請求項１に記載の細胞培養器具であって、
前記第１のスリットは、前記高さ方向に沿って前記外筒全体に形成されている
細胞培養器具。 The cell culture instrument according to claim 1,
The first slit is formed in the entire outer cylinder along the height direction.

請求項１または２に記載の細胞培養器具であって、
前記外筒は、前記第１の内筒の両端部のうちの前記一方の端部により近い側の端部において、該端部の開口を跨ぐように該開口の外周の異なる位置間を連結した第１の把持部を備える
細胞培養器具。 The cell culture instrument according to claim 1 or 2,
The outer cylinder is connected between different positions on the outer periphery of the opening so as to straddle the opening of the end at the end closer to the one end of the both ends of the first inner cylinder. A cell culture instrument comprising a first grip.

請求項１から３のうちのいずれか１項に記載の細胞培養器具であって、さらに、
前記第１の内筒と高さ方向が同じである筒状に形成されて、前記第１の内筒の前記一方の端部側で前記細胞培養用膜を介して前記第１の内筒と接するように前記外筒内に収容される第２の内筒を備える
細胞培養器具。 The cell culture instrument according to any one of claims 1 to 3, further comprising:
The first inner cylinder is formed in a cylindrical shape having the same height direction as the first inner cylinder via the cell culture membrane on the one end side of the first inner cylinder. A cell culture instrument comprising a second inner cylinder accommodated in the outer cylinder so as to come into contact therewith.

請求項４に記載の細胞培養器具であって、
前記第２の内筒は、前記高さ方向の力を加えることにより、前記外筒内で前記高さ方向にスライド可能であって、スライドさせた位置にて保持可能である
細胞培養器具。 The cell culture instrument according to claim 4,
The second inner cylinder is slidable in the height direction within the outer cylinder by applying a force in the height direction, and can be held at the slid position.

請求項１に記載の細胞培養器具であって、
前記外筒に形成される前記第１のスリットは、前記第１の内筒の前記一方の端部と前記外筒とが重なる位置から、前記第１の内筒から高さ方向に離間する方向に向かって延びるように形成されている
細胞培養器具。 The cell culture instrument according to claim 1,
The first slit formed in the outer cylinder is a direction away from the first inner cylinder in a height direction from a position where the one end portion of the first inner cylinder and the outer cylinder overlap. A cell culture instrument formed to extend toward the cell.

請求項１から６のうちのいずれか１項に記載の細胞培養器具であって、
前記第１の内筒は、該第１の内筒の両端部のうちの他方の端部において、該他方の端部の開口を跨ぐように該他方の端部の開口の外周の異なる位置間を連結した第２の把持部を備える
細胞培養器具。 The cell culture instrument according to any one of claims 1 to 6,
The first inner cylinder is located between different positions of the outer periphery of the opening of the other end so as to straddle the opening of the other end of the other end of the first inner cylinder. A cell culture instrument provided with the 2nd holding part which connected.

請求項３に記載の細胞培養器具であって、
前記第１の把持部は、前記第１の内筒の軸線上付近で交わる２本の線分状に形成されている
細胞培養器具。 The cell culture instrument according to claim 3,
The cell culture instrument, wherein the first gripping portion is formed in two line segments that intersect in the vicinity of the axis of the first inner cylinder.

請求項７に記載の細胞培養器具であって、
前記第２の把持部は、前記第１の内筒の軸線上付近で交わる２本の線分状に形成されている
細胞培養器具。 The cell culture instrument according to claim 7,
The cell culture instrument, wherein the second grip portion is formed in two line segments that intersect in the vicinity of the axis of the first inner cylinder.

請求項６に記載の細胞培養器具であって、
前記第１の内筒には、
前記外筒に設けた前記第１のスリットと重なる位置において、前記一方の端部から前記高さ方向に延びる第２のスリットと、
前記外筒に設けた前記第１のスリットと重なる位置であって、前記第１の内筒の前記一方の端部における前記第２のスリットが開口する位置において、前記第１の内筒の径方向外側に向かって前記第１のスリットを経由して前記外筒の外側に延びる一対の膜支持部と、
が設けられている細胞培養器具。 The cell culture instrument according to claim 6, wherein
In the first inner cylinder,
A second slit extending in the height direction from the one end at a position overlapping the first slit provided in the outer cylinder;
The diameter of the first inner cylinder at a position that overlaps the first slit provided in the outer cylinder and at which the second slit at the one end of the first inner cylinder opens. A pair of membrane support portions extending outward of the outer cylinder via the first slit toward the outside in the direction;
A cell culture instrument provided with

請求項１から１０のうちのいずれか１項に記載の細胞培養器具であって、
前記第１の内筒は、該第１の内筒における前記一方の端部近傍の外側面であって、前記細胞培養用膜と接する位置に、凹凸形状が設けられている
細胞培養器具。 The cell culture instrument according to any one of claims 1 to 10,
The first inner cylinder is an outer surface in the vicinity of the one end portion of the first inner cylinder, and an uneven shape is provided at a position in contact with the cell culture membrane.