JPH0568229B2 - - Google Patents
Info
- Publication number
- JPH0568229B2 JPH0568229B2 JP62137959A JP13795987A JPH0568229B2 JP H0568229 B2 JPH0568229 B2 JP H0568229B2 JP 62137959 A JP62137959 A JP 62137959A JP 13795987 A JP13795987 A JP 13795987A JP H0568229 B2 JPH0568229 B2 JP H0568229B2
- Authority
- JP
- Japan
- Prior art keywords
- cells
- hair
- hair bulb
- bulb
- culture
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
- 210000004209 hair Anatomy 0.000 claims description 81
- 210000004027 cell Anatomy 0.000 claims description 54
- 230000002500 effect on skin Effects 0.000 claims description 12
- 210000002950 fibroblast Anatomy 0.000 claims description 11
- 238000012258 culturing Methods 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 10
- 102000004142 Trypsin Human genes 0.000 claims description 9
- 108090000631 Trypsin Proteins 0.000 claims description 9
- 239000012588 trypsin Substances 0.000 claims description 9
- 102000029816 Collagenase Human genes 0.000 claims description 8
- 108060005980 Collagenase Proteins 0.000 claims description 8
- 229960002424 collagenase Drugs 0.000 claims description 8
- 108010007093 dispase Proteins 0.000 claims description 8
- 210000003491 skin Anatomy 0.000 claims description 8
- 210000001519 tissue Anatomy 0.000 claims description 8
- 210000002615 epidermis Anatomy 0.000 claims description 6
- 102000004190 Enzymes Human genes 0.000 claims description 4
- 108090000790 Enzymes Proteins 0.000 claims description 4
- 229940088598 enzyme Drugs 0.000 claims description 4
- 238000005119 centrifugation Methods 0.000 claims description 2
- 210000004207 dermis Anatomy 0.000 description 9
- 210000004694 pigment cell Anatomy 0.000 description 8
- 210000001339 epidermal cell Anatomy 0.000 description 7
- 239000011159 matrix material Substances 0.000 description 7
- 210000004927 skin cell Anatomy 0.000 description 7
- 239000000725 suspension Substances 0.000 description 5
- 238000001000 micrograph Methods 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 3
- 241000699666 Mus <mouse, genus> Species 0.000 description 3
- 239000012091 fetal bovine serum Substances 0.000 description 3
- 230000003780 keratinization Effects 0.000 description 3
- 239000010410 layer Substances 0.000 description 3
- 230000035945 sensitivity Effects 0.000 description 3
- 201000004384 Alopecia Diseases 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 102000009016 Cholera Toxin Human genes 0.000 description 2
- 108010049048 Cholera Toxin Proteins 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 210000004748 cultured cell Anatomy 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 230000003779 hair growth Effects 0.000 description 2
- 208000024963 hair loss Diseases 0.000 description 2
- 230000003676 hair loss Effects 0.000 description 2
- 206010033675 panniculitis Diseases 0.000 description 2
- 238000012827 research and development Methods 0.000 description 2
- 210000004918 root sheath Anatomy 0.000 description 2
- 210000004304 subcutaneous tissue Anatomy 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 241000700198 Cavia Species 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 102000011782 Keratins Human genes 0.000 description 1
- 108010076876 Keratins Proteins 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 210000003969 blast cell Anatomy 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 238000004141 dimensional analysis Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 231100000640 hair analysis Toxicity 0.000 description 1
- 210000002768 hair cell Anatomy 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 210000001208 inner root sheath cell Anatomy 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 238000005191 phase separation Methods 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
Description
【発明の詳細な説明】
産業上の利用分野
本発明は、マウス、ラツト、モルモツト、ある
いは人の毛球部細胞の培養方法に関する。DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application The present invention relates to a method for culturing hair bulb cells of mice, rats, guinea pigs, or humans.
従来の技術および問題点
毛球部は、毛母細胞を含み発毛の根源をなすも
のである。すなわち、毛母細胞は、内根鞘、小
皮、皮質、髄質に分化する。このような毛球部の
細胞の培養を行うことは、育毛剤の開発研究、並
びに毛の基礎研究のために極めて重要である。Prior Art and Problems The hair bulb contains hair matrix cells and is the source of hair growth. That is, hair matrix cells differentiate into the inner root sheath, cuticle, cortex, and medulla. Cultivating such cells of the hair bulb is extremely important for research and development of hair restorers as well as basic research on hair.
従来、毛の細胞培養方法に関しては、いくつか
の報告がある。しかしながら、これらの培養細胞
は、すでに分化した後の毛根鞘由来の細胞にかか
るものであつて、いわゆる多岐に分化する前の毛
母細胞を含む毛球部の細胞を培養した例はない。 There have been several reports regarding hair cell culture methods. However, these cultured cells involve cells derived from the hair root sheath that have already differentiated, and there is no example of culturing cells of the hair bulb including hair matrix cells that have not yet differentiated into various types.
本発明は、増殖、分化能力を有し、角化状態に
至る毛球部を培養する方法を提供することを目的
とする。 An object of the present invention is to provide a method for culturing hair bulbs that have the ability to proliferate and differentiate and reach a keratinized state.
問題点を解決するための手段
本発明は、表皮および／または真皮細胞が付着
したままの毛球部を含む皮膚を酵素処理して、表
皮および／または真皮細胞、並びに毛球部からな
る懸濁液をえ、該懸濁液より毛細部を分離して組
織培養を行うことを特徴とする毛球部細胞の培養
方法を提供するものである。Means for Solving the Problems The present invention involves enzymatically treating the skin containing the hair bulb to which the epidermis and/or dermal cells are attached, thereby creating a suspension consisting of the epidermis and/or dermal cells and the hair bulb. The present invention provides a method for culturing hair bulb cells, which comprises obtaining a suspension, separating hair particles from the suspension, and performing tissue culture.
本発明によれば、損傷することなく多数の毛球
部細胞が容易に得られる。本発明にて得られた毛
球部の細胞は、増殖、分化能力を持ち、最終的に
は角化状態の特徴を呈する。すなわち、生体内に
おいて生ずる生理学的現象が、in vitroのシヤー
レ内にて生ずる、
なお、毛球部の細胞を培養した場合には、僅か
ながら真皮線維芽細胞が混入する。しかしなが
ら、このような混入真皮線維芽細胞は、細胞をシ
ヤーレに植え込んだ後、毛球部の細胞とのトリプ
シン感受性の差を利用してとり除くことができ
る。したがつて、本発明により毛球部の細胞の純
粋培養が可能となる。 According to the present invention, a large number of hair bulb cells can be easily obtained without being damaged. The cells of the hair bulb obtained according to the present invention have the ability to proliferate and differentiate, and eventually exhibit the characteristics of a keratinized state. In other words, physiological phenomena that occur in a living body occur in an in vitro shear cell. Note that when hair bulb cells are cultured, a small amount of dermal fibroblasts are mixed in. However, such contaminating dermal fibroblasts can be removed by using the difference in trypsin sensitivity from cells in the hair bulb after the cells are implanted in the shear. Therefore, the present invention enables pure culture of hair bulb cells.
これに対し、従来、分化前の毛球部の細胞培養
が行えなかつた原因は、細胞を得る方法が毛の抜
去、もしくは皮下織、真皮をメスで切り離すとい
う方法であるため、毛球部に損傷を与えているか
らと考えられる。真皮と毛、あるいは表皮と毛、
相互間の結合はかなり強く、したがつて、抜去に
よる細胞の採取はそれら相互間の結合力に抗して
行うもので毛球部細胞は損傷を受けたり、あるい
は皮膚内に残留する。 On the other hand, the reason why it has not been possible to culture cells in the hair bulb before differentiation is that the method of obtaining cells is to remove the hair or cut off the subcutaneous tissue and dermis with a scalpel. This is probably due to damage. dermis and hair, or epidermis and hair,
The bonds between hair bulbs are quite strong, and therefore, when cells are removed by removal, the hair bulb cells are damaged or remain in the skin.
本発明者らが行つた従来法による細胞の採取、
培養試験においても、600例の抜去した毛の培養
において、成功例はわずか２例にすぎず、また、
これら成功例にあつても、毛根鞘の細胞が遊走、
増殖するのみであり、毛球部の細胞はすべて死細
胞であり増殖可能な毛球部の細胞を得ることはで
きなかつた。 Collection of cells by the conventional method performed by the present inventors,
In culture tests, out of 600 cases of culture of removed hair, only 2 cases were successful.
Even in these successful cases, the cells of the hair root sheath migrate,
All the cells in the hair bulb were dead cells, and it was not possible to obtain cells in the hair bulb that could proliferate.
本発明によれば、毛の色素細胞を多数得ること
が可能である。毛球部を構成する重要な細胞の１
つである毛の色素細胞を培養した例は従来なく、
毛母色素細胞と毛母細胞の関係を調べることは、
脱毛および白髪化の両面から毛の老化現象の解明
に有用である。 According to the present invention, it is possible to obtain a large number of hair pigment cells. One of the important cells that make up the hair bulb
There has never been an example of culturing hair pigment cells, which are
To investigate the relationship between hair matrix pigment cells and hair matrix cells,
It is useful for elucidating the phenomenon of hair aging in terms of both hair loss and graying.
本発明にて培養に用いられる毛球部を含む毛髪
サンプルは、表紙細胞、または真皮細胞、あるい
はこれらの両方が付着したままのものを用いる。
該サンプルは、毛球部を含む皮膚を切除して得ら
れた。 The hair sample containing the hair bulb used for culture in the present invention is one to which cover cells, dermal cells, or both remain attached.
The sample was obtained by excising the skin including the hair bulb.
つぎに、かかる試料より毛球部の細胞を得るに
は、表皮細胞と毛、真皮細胞と毛の各々相互間の
結合を酵素処理により取り除くことにより行なわ
れる。このような酵素処理に用いられる酵素とし
ては、例えば、コラゲナーゼ、デイスパーゼ、ト
リプシンなどが挙げらる。コラゲナーゼおよびデ
イスパーゼはCa2+およびMg2+を含有する培養液
を、またトリプシンはPBS（−）を溶液として用
いるのが好ましく、酵素溶液の濃度は、通常、デ
イスパーゼ500〜10000U／ml、コラゲナーゼ0.25
％、トリプシン0.15〜0.25％であるのが好まし
い。 Next, hair bulb cells can be obtained from such a sample by removing the bonds between epidermal cells and hair and between dermal cells and hair by enzymatic treatment. Examples of enzymes used in such enzymatic treatment include collagenase, dispase, and trypsin. It is preferable to use a culture solution containing Ca 2+ and Mg 2+ for collagenase and dispase, and to use PBS (-) as a solution for trypsin. The concentration of the enzyme solution is usually 500 to 10,000 U/ml for dispase and 0.25 U/ml for collagenase.
%, preferably trypsin 0.15-0.25%.
つぎに、かかる酵素処理により得られた表紙細
胞、真皮細胞および毛球部細胞がばらばらになつ
た懸濁液を遠心分離などを用いて相分離するか、
あるいはパーコールの密度勾配を用いて分離し純
粋の毛球部を得る。 Next, the suspension in which the cover cells, dermal cells, and hair bulb cells obtained by the enzyme treatment are separated is subjected to phase separation using centrifugation or the like, or
Alternatively, a pure hair bulb is obtained by separating using a Percoll density gradient.
このようにして得られた毛球部を組織培養す
る。組織培養を行うには、牛胎児血清を添加した
培養液を使用し、37℃、５％CO2下、インキユベ
ーター内で培養する。 The hair bulb thus obtained is tissue cultured. To perform tissue culture, a culture medium supplemented with fetal bovine serum is used, and the tissue is cultured in an incubator at 37° C. and 5% CO 2 .
なお、毛球部細胞の培養を行つた後、さらにト
リプシンの感受性の差に基づき、真皮線維芽細胞
を除去し、毛球部の細胞のみを単離するのが好ま
しい。 After culturing the hair bulb cells, it is preferable to remove the dermal fibroblasts and isolate only the hair bulb cells based on the difference in sensitivity to trypsin.
実施例
つぎに、本発明を実施例によりさらに具体的に
説明する。Examples Next, the present invention will be explained in more detail with reference to examples.
(a) 毛球部を得る方法
マウスをクロロホルムでと殺し、70％エタノ
ールで皮膚を消毒した。皮膚を採取し、イーグ
ル培養液（Eagle′s Minimum Essential
Medium：以下、MEMと略す）で洗浄後、皮
下織、血管をていねいにメスで取り除いた。こ
の皮膚を約９mm2の大きさに細断し、500もしく
は1000U／mlのデイスパーゼ溶液（５％牛胎児
血清（FBS）・MEM内）に浸積し、一昼夜４
℃下で静置した。デイスパーゼ処理後、ピンセ
ツトでていねいに表皮を剥離除去して真皮を得
た。得られた真皮をPBS（−）に浸積し、デイ
スパーゼを失活させた。(a) Method for obtaining hair bulbs Mice were killed with chloroform, and the skin was disinfected with 70% ethanol. The skin was collected and cultured in Eagle's Minimum Essential
After washing with medium (hereinafter abbreviated as MEM), the subcutaneous tissue and blood vessels were carefully removed with a scalpel. This skin was cut into pieces approximately 9 mm2 in size, soaked in 500 or 1000 U/ml dispase solution (5% fetal bovine serum (FBS) in MEM), and incubated overnight for 4 days.
It was left standing at ℃. After dispase treatment, the epidermis was carefully peeled off and removed using forceps to obtain the dermis. The obtained dermis was immersed in PBS(-) to inactivate dispase.
つぎに、この真皮を0.25％コラゲナーゼ
（MEM内）により処理した。コラゲナーゼ処
理時間は、用いた皮膚の種属・日令により変化
させてよいが、いずれも37℃下で行う。コラゲ
ナーゼ溶液内で軽く混ぜるだけで真皮が完全に
懸濁状態になる。すなわち、真皮が組織の原形
をとどめず、細胞１つ１つになる状態までコラ
ゲナーゼ処理を行う。この状態では、真皮細胞
の中に丸い球形の毛球部が浮遊している。 This dermis was then treated with 0.25% collagenase (in MEM). The collagenase treatment time may vary depending on the species and age of the skin used, but it is performed at 37°C. The dermis is completely suspended in the collagenase solution by simply mixing it briefly. That is, the collagenase treatment is performed until the dermis does not retain its original tissue shape but becomes individual cells. In this state, round hair bulbs are floating within the dermal cells.
つぎに、この懸濁液を1000rpm、５分間遠心
して毛球部の層を得た。得られた層をPBS
（−）で３回洗浄することにより、混入した真
皮線維芽細胞をできる限り取り除いた。得られ
た層には真皮線維芽細胞は殆ど存在せずに毛球
部が優勢の状態で浮遊している。（第１図参照）
つぎに、得られた毛球部を10％FBS−MEM
（抗生物質および10ng／mlのコレラトキシンを
含む）を加え、軽く攪拌し、組織培養用シヤー
レに植え込み、５％CO2インキユベーター内
（37℃）で培養したところ、多くの毛球部の細
胞が遊走し増殖した。（第２図参照）
(b) 毛球部単離培養方法
上記の方法によりほとんど真皮を取り除くこ
とができるが、なお僅かながら真皮線維芽細胞
が混入する。培養後、真皮線維芽細胞を除去
し、毛球部の細胞のみを単離するには、毛球部
の細胞と真皮細胞の両細胞のトリプシン感受性
の差を利用する。 Next, this suspension was centrifuged at 1000 rpm for 5 minutes to obtain a hair bulb layer. PBS the resulting layer
By washing with (-) three times, as much as possible of the contaminated dermal fibroblasts was removed. In the resulting layer, there are almost no dermal fibroblasts, and hair bulbs are predominant and floating. (See Figure 1) Next, the obtained hair bulb was mixed with 10% FBS-MEM.
(containing antibiotics and 10 ng/ml of cholera toxin), stirred gently, implanted in a tissue culture shear dish, and cultured in a 5% CO 2 incubator (37°C). Cells migrated and proliferated. (See Figure 2) (b) Hair bulb isolation and culture method Although most of the dermis can be removed by the above method, a small amount of dermal fibroblasts will still be mixed in. After culturing, the dermal fibroblasts are removed and only the hair bulb cells are isolated by utilizing the difference in trypsin sensitivity between the hair bulb cells and the dermal cells.
まずPBS（−）で細胞をよく洗浄し、さらに
0.02％EDTAで洗う。その後、0.15％トリプシ
ン（37℃、５分間）処置をする。これらの一連
の操作を施すと、毛球部の細胞はシヤーレに付
着するが、混入した真皮線維芽細胞は浮遊す
る。また、10ng／mlのコレラトキシンを添加
することにより、真皮線維芽細胞の増殖を抑制
することができる。培養の結果、真皮線維芽細
胞は除去され毛球部の細胞の純粋培養を行うこ
とができた。（第３図参照）
培養されたこれらの細胞は、つぎの方法により
毛球部の細胞であることが確認された。 First, wash the cells thoroughly with PBS (-), and then
Wash with 0.02% EDTA. Then, treat with 0.15% trypsin (37°C, 5 minutes). When a series of these operations is performed, the cells of the hair bulb adhere to the shear, but the contaminated dermal fibroblasts float. Further, by adding 10 ng/ml of cholera toxin, proliferation of dermal fibroblasts can be suppressed. As a result of the culture, dermal fibroblasts were removed and pure culture of cells from the hair bulb region could be performed. (See Figure 3) These cultured cells were confirmed to be hair bulb cells by the following method.
(1) 毛球部の細胞と表皮細胞のケラチンパターン
を等電点、分子量により二次元解析すると、
各々のケラチスポツトに相違がある。(1) Two-dimensional analysis of keratin patterns in hair bulb cells and epidermal cells using isoelectric point and molecular weight reveals that
Each keratis spot is different.
(2) 通常マウス表皮には色素細胞は存在しない。
従つて、表皮細胞を培養すると色素細胞が共存
していることはない。一方、マウス毛球部には
多数の色素細胞が存在する。(2) Normally, there are no pigment cells in the mouse epidermis.
Therefore, when epidermal cells are cultured, pigment cells do not coexist. On the other hand, there are many pigment cells in the mouse hair bulb.
本発明によりマウス毛球部の細胞を単離培養
すると、多数の毛母色素細胞が共存している。
（第４図参照）
(3) 表皮細胞を通常培養液のCa濃度で培養する
と、角化が起こり、毛球部の細胞も同様に角化
が起こる。しかし、両者の角化の状態には相違
がある。すなわち、表皮細胞はシート状に角化
するが、毛球部の細胞は縦に長く線維状に角化
する。 When cells of the mouse hair bulb are isolated and cultured according to the present invention, a large number of hair matrix pigment cells coexist.
(See Figure 4) (3) When epidermal cells are cultured at the Ca concentration of a normal culture medium, keratinization occurs, and cells in the hair bulb also undergo keratinization. However, there is a difference in the state of keratinization between the two. That is, epidermal cells are keratinized in a sheet-like manner, whereas cells in the hair bulb are keratinized in a longitudinally long fibrous form.
(4) シヤーレに対する付着率について、表皮細胞
と毛球部の細胞の間で相違がある。表皮細胞は
組織培養用プラスチツクシヤーレでも付着する
率は悪く、ガラスシヤーレ並びにバクテリア用
シヤーレにはほとんど付着することはない。(4) There is a difference between epidermal cells and cells in the hair bulb regarding the adhesion rate to shear. The adhesion rate of epidermal cells to plastic shears for tissue culture is poor, and they almost never adhere to glass shears or bacterial shears.
本発明により得られた毛球部のシヤーレに対
する付着率は良く、ガラスシヤーレ並びにバク
テリア用シヤーレでも付着し得る。 The hair bulb obtained according to the present invention has a good adhesion rate to shear, and can adhere to glass shear as well as bacterial shear.
本発明の方法により培養された毛球部の細胞
は、育毛剤開発研究や、多方面にわたる毛の基礎
研究に使用しうる。したがつて、毛の発生に関与
する因子をとらえ、永点未解決のままであつた脱
毛の機構の解明の手だてとなる。 Hair bulb cells cultured by the method of the present invention can be used for research on the development of hair growth agents and basic research on hair in a wide range of fields. Therefore, it will help us understand the factors involved in hair development and elucidate the mechanism of hair loss, which has remained unresolved for a long time.
発明の効果
本発明によれば、増殖、分化能力を有した角化
状態に至る毛球部の培養が可能となり、育毛剤の
開発研究、並びに毛の基礎研究に利用することが
できる。Effects of the Invention According to the present invention, it is possible to culture the hair bulb to a keratinized state with the ability to proliferate and differentiate, and it can be used for research and development of hair restorers as well as basic research on hair.
第１図は真皮より得られた優勢の状態で浮遊す
る毛球部の形態を示す顕微鏡写真、第２図は遊
走、増殖する毛球部細胞の形態を示す顕微鏡写
真、第３図は真皮線維芽細胞が除去され単層純粋
培養された毛球部細胞の形態を示す顕微鏡写真、
第４図は毛母色素細胞の形態を示す顕微鏡写真で
ある。
Figure 1 is a micrograph showing the morphology of a floating hair bulb obtained from the dermis, Figure 2 is a micrograph showing the morphology of migrating and proliferating hair bulb cells, and Figure 3 is a dermal fiber. A micrograph showing the morphology of hair bulb cells that have been cultured in a pure monolayer with blast cells removed;
FIG. 4 is a micrograph showing the morphology of hair matrix pigment cells.
Claims (1)
処理し、表皮を剥離除去した後、コラーゲナー
ゼ、デイスパーゼおよびトリプシンから選ばれた
少なくとも１種の酵素を用いて毛球部を他の細胞
から解離させ、ついで遠心分離法により毛球部細
胞を分離して培養することを特徴とする毛球部細
胞の培養方法。 ２ 分離および培養して得られた毛球部細胞をさ
らに、トリプシン処理して真皮線維芽細胞を除去
する前記特許請求の範囲第１項記載の方法。[Claims] 1. After treating skin tissue containing hair bulb cells with dispase and exfoliating the epidermis, the hair bulb is treated with at least one enzyme selected from collagenase, dispase, and trypsin. 1. A method for culturing hair bulb cells, which comprises dissociating hair bulb cells from other cells, and then separating and culturing hair bulb cells by centrifugation. 2. The method according to claim 1, wherein the hair bulb cells obtained by separating and culturing are further treated with trypsin to remove dermal fibroblasts.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62137959A JPS63301785A (en) | 1987-06-01 | 1987-06-01 | Culture of hair-bulb cell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP62137959A JPS63301785A (en) | 1987-06-01 | 1987-06-01 | Culture of hair-bulb cell |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS63301785A JPS63301785A (en) | 1988-12-08 |
| JPH0568229B2 true JPH0568229B2 (en) | 1993-09-28 |
Family
ID=15210724
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP62137959A Granted JPS63301785A (en) | 1987-06-01 | 1987-06-01 | Culture of hair-bulb cell |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS63301785A (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2006055106A (en) * | 2004-08-23 | 2006-03-02 | National Institute Of Advanced Industrial & Technology | Method for culturing mesenchymal stem cell derived from human bone marrow by using human serum culture medium |
-
1987
- 1987-06-01 JP JP62137959A patent/JPS63301785A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS63301785A (en) | 1988-12-08 |
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