CN108315258A - Increase cell culture roller bottle and its application of cell culture area - Google Patents
Increase cell culture roller bottle and its application of cell culture area Download PDFInfo
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- CN108315258A CN108315258A CN201810445820.XA CN201810445820A CN108315258A CN 108315258 A CN108315258 A CN 108315258A CN 201810445820 A CN201810445820 A CN 201810445820A CN 108315258 A CN108315258 A CN 108315258A
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- 238000004113 cell culture Methods 0.000 title claims abstract description 56
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 7
- 210000001178 neural stem cell Anatomy 0.000 claims description 7
- 239000012530 fluid Substances 0.000 claims description 4
- 239000004793 Polystyrene Substances 0.000 claims description 3
- 239000004698 Polyethylene Substances 0.000 claims description 2
- 239000000463 material Substances 0.000 claims description 2
- -1 polyethylene Polymers 0.000 claims description 2
- 229920000573 polyethylene Polymers 0.000 claims description 2
- 229920002223 polystyrene Polymers 0.000 claims description 2
- 210000000130 stem cell Anatomy 0.000 claims description 2
- 150000003503 terephthalic acid derivatives Chemical class 0.000 claims description 2
- 230000004083 survival effect Effects 0.000 abstract description 5
- 210000004027 cell Anatomy 0.000 description 22
- 230000010261 cell growth Effects 0.000 description 5
- 230000001464 adherent effect Effects 0.000 description 4
- 230000003833 cell viability Effects 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 239000004033 plastic Substances 0.000 description 4
- 229920003023 plastic Polymers 0.000 description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 150000001336 alkenes Chemical class 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 238000003306 harvesting Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 229920006389 polyphenyl polymer Polymers 0.000 description 2
- 239000002356 single layer Substances 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 244000261422 Lysimachia clethroides Species 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 206010044074 Torticollis Diseases 0.000 description 1
- GLNADSQYFUSGOU-GPTZEZBUSA-J Trypan blue Chemical compound [Na+].[Na+].[Na+].[Na+].C1=C(S([O-])(=O)=O)C=C2C=C(S([O-])(=O)=O)C(/N=N/C3=CC=C(C=C3C)C=3C=C(C(=CC=3)\N=N\C=3C(=CC4=CC(=CC(N)=C4C=3O)S([O-])(=O)=O)S([O-])(=O)=O)C)=C(O)C2=C1N GLNADSQYFUSGOU-GPTZEZBUSA-J 0.000 description 1
- 238000004115 adherent culture Methods 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000005034 decoration Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 238000005213 imbibition Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 208000018197 inherited torticollis Diseases 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/08—Flask, bottle or test tube
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/34—Internal compartments or partitions
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0618—Cells of the nervous system
- C12N5/0623—Stem cells
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- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Zoology (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Clinical Laboratory Science (AREA)
- Sustainable Development (AREA)
- Developmental Biology & Embryology (AREA)
- Neurology (AREA)
- Neurosurgery (AREA)
- Cell Biology (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
A kind of cell culture roller bottle increasing cell culture area, including bottle and bottleneck.Bottle includes bottom of bottle, bottle inner wall, bottle inner cavity and bottle top, and bottle inner cavity is arranged at least 1 deflector, and the week of deflector with bottom of bottle, bottle inner wall or bottle top along being connected at least one；Water conservancy diversion plate thickness is 1 millimeter~4 millimeters, and length is 1/2 or more of bottle depth of inner cavity, and width is less than the radius of bottle, and plate face is sprawled along the radial direction of bottle.The inner cavity of bottleneck is connected to bottle intracavity.The cell culture roller bottle provided by the invention for increasing cell culture area significantly improves living cells culture quantity, and cell survival rate does not reduce.
Description
Technical field
The present invention relates to a kind of container more particularly to a kind of containers of culture cell, to increase cell culture area.
Background technology
Small-scale zooblast organizes the container of in vitro culture mainly to use Tissue Culture Flask.Commodity Tissue Culture Flask
Material is mainly two kinds of glass and plastics, and the bottom shape of bottle has square, rectangle, triangle, trapezoidal etc.；Cell is adherent
Culture area is 25cm2~175cm2, culture bottle volume has 5ml to 500ml.Since carbon dioxide training is placed in the more accumbency of cell bottle
Case is supported, in order to which imbibition or harvest are convenient, bottleneck pattern also there are many design, there is straight neck, torticollis, angle neck, triangle, rectangle
Deng.Most cells culture vessel is mostly single layer vessel at present, such as conventional capacity 50ml (culture area 25cm2) and capacity
250ml (culture area 75cm2) plastic cell culture bottle.
When carrying out mass cell culture for the zooblast of adherent growth, there are many strategies.One is using glass
Or stainless steel bioreactor, cell need attaching carrier circular disc to be grown, need stirring to ventilate, this kind of cell culture mesh
It is main with collect cell secretion product based on, as monoclonal antibody produce.It is for second the culture of the bigger specification of customization
Bottle increases the adherent area in cell bottle bottom, with Thermo Fischer Scient Inc. (Thermo Fisher
ScientificTMNuncTM) and Corning Incorporated (Corning Incoporated) be representative cell factory system, using more
Confluent monolayer cells bottle is superimposed.The third is the cell culture roller bottle using traditional tubular.
In actual production, the cell culture roller bottle of tubular is used for expanding adherent or suspension growth cell, and there are many excellent
Gesture.Benefit using roller bottle culture is at low cost, technology maturation, easy to operate.Increase roller bottle quantity in production as needed, increases
The culture amount of refinement born of the same parents.
Invention content
It is an object of the present invention to provide a kind of cell culture roller bottles increasing cell culture area, improve adherent thin
Born of the same parents' culture amount.
It is another object of the present invention to provide a kind of cell culture roller bottles increasing cell culture area, are suitable for dry
The amplification cultivation of cell and other cell types.
A kind of cell culture roller bottle increasing cell culture area, including bottle and bottleneck, bottle include bottom of bottle, in bottle
Wall, bottle inner cavity and bottle top.At least 1 deflector is arranged in bottle inner cavity, at least one the week of deflector along and bottom of bottle, bottle inner wall or bottle
Top is connected.
Bottleneck inner cavity is connected to bottle intracavity.
Deflector is made of polystyrene, polyethylene or poly terephthalic acid class plastics etc., and thickness is 1 millimeter~4 millis
Rice, length are 1/2 or more of bottle depth of inner cavity, especially the 80%~90% of bottle depth of inner cavity, and width is less than the radius of bottle,
Its plate face is sprawled along the radial direction of bottle.
The side of deflector connects with bottle internal chamber wall, and both ends push up to form gap with bottom of bottle and bottle respectively, the width in gap
It is the 5%~10% of bottle depth of inner cavity so that between the quasi- number of fluid in bottle everywhere is 2500~3000.
The plate face of deflector preferentially selects fanning strip, to generate the stable quasi- number of fluid.
Baffle quantity is 2 or more, such as：But 2,4,6,8 and 10 etc. are not limited to, in pairs, the diameter along bottle is set
It sets, is distributed in the both sides of bottle axial direction, preferentially select 4 deflectors.
Roller bottle provided by the invention is cultivated applied to stem cell, especially in the culture of neural stem cell, can be significantly improved
Cell viability, cell survival rate also significantly improve.
The advantageous effect that technical solution of the present invention is realized：
The cell culture roller bottle provided by the invention for increasing cell culture area is on the basis of existing roller bottle, along bottle
Radial direction, bottle intracavity increase deflector, especially with sector deflector, realize cell in solid space grow,
Cell growth area is not only expanded, cell culturing space is saved, significantly improves cell culture amount, improves cell culture effect
Rate.
The cell culture roller bottle provided by the invention for increasing cell culture area significantly improves cell viability, cell survival
Rate also significantly improves.
Description of the drawings
Fig. 1 is the structural schematic diagram for one embodiment of cell culture roller bottle that the present invention increases cell culture area；
Fig. 2 is the structural schematic diagram for another angle of cell culture roller bottle for increasing cell culture area shown in Fig. 1.
Specific implementation mode
Below in conjunction with attached drawing detailed description of the present invention technical solution.The embodiment of the present invention is only to illustrate the skill of the present invention
Art scheme and it is unrestricted, although being described the invention in detail with reference to preferred embodiment, those skilled in the art
It should be appreciated that can be modified or replaced equivalently to the technical solution of invention, without departing from the essence of technical solution of the present invention
God and range, should all cover in scope of the presently claimed invention.
Fig. 1 is the structural schematic diagram for one embodiment of cell culture roller bottle that the present invention increases cell culture area, and Fig. 2 is figure
Increase the structural schematic diagram of another angle of cell culture roller bottle of cell culture area shown in 1.As depicted in figs. 1 and 2, this implementation
The cell culture roller bottle of the increase cell culture area of example, includes the plastic bottle body 1 and bottleneck 2 of integrally connected.Bottle 1 includes bottle
Bottom, bottle inner wall, bottle inner cavity and bottle top.At least 1 deflector is arranged in bottle intracavity, at least one the week of deflector along with bottom of bottle,
Bottle inner wall or bottle top are connected.
In the present embodiment, bottle inner wall is uniformly distributed 4 deflectors 3, and in pairs, the diameter along bottle is arranged, and is distributed in
The both sides of bottle axial direction.Each water conservancy diversion plate thickness is 1 millimeter~4 millimeters, and length is 1/2 or more of bottle depth of inner cavity, especially
The 80%~90% of bottle depth of inner cavity, width is less than the radius of bottle, and plate face is sprawled along the radial direction of bottle.Each deflector one
Side connects with bottle intracavity wall, and gap is arranged in both ends with bottom of bottle and bottle top respectively, and the width in gap is bottle depth of inner cavity
5%~10% so that between the quasi- number of fluid in bottle everywhere is 2500~3000, cultivated cell viability is improved with this,
Cell survival rate significantly improves.
The deflector of the present embodiment also expands cell growth area, saves cell culturing space, significantly improves cell
Culture amount improves cell culture efficiency.
With healthy and free from worry 2000ml (850cm2) for polyphenyl alkene roller bottle size (
850cm2Polystyrene Roller Bottle), outer diameter 117mm, high 271mm, bottleneck outer diameter 45mm.170ml culture mediums
Under, effective cell area of attachment is 850cm2.Area is equivalent to about 11 75cm2 gooseneck square media bottles.
It is fan-shaped by wheel in body using roller bottle provided in this embodiment, 90 degree are spaced, increases by 4 deflectors of symmetry, greatly
Small 60mm × 210mm, since culture plate two sides can provide cell growth face, additional increased cell growth area is
1008cm2, equivalent to increase the cell growth area of a roller bottle, living cells quantity dramatically increases.
10ml is contained to DMEM culture mediums (10% fetal calf serum of about 1,000,000 mouse neural stem cells C17.2
(GIBICO), 5% horse serum (GIBICO) is added to healthy and free from worry 2000ml (850cm2) in polyphenyl alkene roller bottle, add on about 160ml
State DMEM culture mediums.Neural stem cell is after roller bottle culture in 5~7 days, through 0.25% trypsin-EDTA solutions
(Invitrogen) elution harvest neural stem cell.Centrifugation, the neural stem cell after washing are resuspended in 10ml DMEM culture mediums
In, cell survival rate is detected through 0.4% trypan blue (Invitrogen) decoration method, cell viability reaches 95%, cell quantity
About 1,700,000.This show neural stem cell C17.2 can in roller bottle well-grown.
Claims (9)
1. a kind of cell culture roller bottle increasing cell culture area, it is characterised in that including bottle and bottleneck,
The bottle includes bottom of bottle, bottle inner wall, bottle inner cavity and bottle top, and at least 1 deflector is arranged in the bottle inner cavity, at least
The week edge of deflector is connected with the bottom of bottle, the bottle inner wall or the bottle top at one；
The water conservancy diversion plate thickness is 1 millimeter~4 millimeters, and length is 1/2 or more of the bottle depth of inner cavity, and width is less than institute
The radius of bottle is stated, plate face is sprawled along the radial direction of the bottle；
The inner cavity of the bottleneck is connected to the bottle intracavity.
2. the cell culture roller bottle according to claim 1 for increasing cell culture area, it is characterised in that the water conservancy diversion
Plate length is the 80%~90% of the bottle depth of inner cavity.
3. the cell culture roller bottle according to claim 1 for increasing cell culture area, it is characterised in that the water conservancy diversion
Plate quantity is 2 or more, and in pairs, and the diameter along the bottle is arranged, and is distributed in the both sides of the bottle axial direction.
4. the cell culture roller bottle according to claim 1 for increasing cell culture area, it is characterised in that the deflector
Side connect with the bottle internal chamber wall, both ends push up to form gap with the bottom of bottle and the bottle respectively, gap
Width is the 5%~10% of bottle depth of inner cavity.
5. the cell culture roller bottle according to claim 1 for increasing cell culture area, it is characterised in that the water conservancy diversion
The plate face of plate is sector.
6. the cell culture roller bottle according to claim 1 for increasing cell culture area, it is characterised in that the water conservancy diversion
Plate is made of polystyrene, polyethylene or poly terephthalic acid class material.
7. the cell culture roller bottle according to claim 1 for increasing cell culture area, it is characterised in that the bottle
Between the quasi- number of fluid inside everywhere is 2500~3000.
8. application of the cell culture roller bottle according to claim 1 for increasing cell culture area in stem cell is cultivated.
9. cell culture roller bottle the answering in Culture of neural stem cells according to claim 1 for increasing cell culture area
With.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810445820.XA CN108315258A (en) | 2018-05-10 | 2018-05-10 | Increase cell culture roller bottle and its application of cell culture area |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810445820.XA CN108315258A (en) | 2018-05-10 | 2018-05-10 | Increase cell culture roller bottle and its application of cell culture area |
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| Publication Number | Publication Date |
|---|---|
| CN108315258A true CN108315258A (en) | 2018-07-24 |
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| CN201810445820.XA Pending CN108315258A (en) | 2018-05-10 | 2018-05-10 | Increase cell culture roller bottle and its application of cell culture area |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11111470B2 (en) | 2019-02-05 | 2021-09-07 | Corning Incorporated | Packed-bed bioreactor systems and methods of using the same |
| US11118151B2 (en) | 2019-11-05 | 2021-09-14 | Corning Incorporated | Fixed bed bioreactor and methods of using the same |
| US11680237B2 (en) | 2017-12-20 | 2023-06-20 | Univercells Technologies S.A. | Bioreactor and related methods |
| CN116814396A (en) * | 2023-06-29 | 2023-09-29 | 广州沙艾生物科技有限公司 | Stem cell culture device |
| US11827873B2 (en) | 2016-11-09 | 2023-11-28 | Univercells Technologies S.A. | Cell growth matrix |
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Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11827873B2 (en) | 2016-11-09 | 2023-11-28 | Univercells Technologies S.A. | Cell growth matrix |
| US11680237B2 (en) | 2017-12-20 | 2023-06-20 | Univercells Technologies S.A. | Bioreactor and related methods |
| US11111470B2 (en) | 2019-02-05 | 2021-09-07 | Corning Incorporated | Packed-bed bioreactor systems and methods of using the same |
| US11401493B2 (en) | 2019-02-05 | 2022-08-02 | Corning Incorporated | Packed-bed bioreactor systems and methods of using the same |
| US11434460B2 (en) | 2019-02-05 | 2022-09-06 | Corning Incorporated | Woven cell culture substrates, bioreactor systems using the same, and related methods |
| US11639489B2 (en) | 2019-02-05 | 2023-05-02 | Corning Incorporated | Packed-bed bioreactor systems and methods of using the same |
| US11661576B2 (en) | 2019-02-05 | 2023-05-30 | Corning Incorporated | Packed-bed bioreactor systems and methods of using the same |
| US11692161B2 (en) | 2019-02-05 | 2023-07-04 | Corning Incorporated | Packed-bed bioreactor systems and methods of using the same |
| US11920117B2 (en) | 2019-02-05 | 2024-03-05 | Corning Incorporated | Woven cell culture substrates, bioreactor systems using the same, and related methods |
| US11118151B2 (en) | 2019-11-05 | 2021-09-14 | Corning Incorporated | Fixed bed bioreactor and methods of using the same |
| US11795426B2 (en) | 2019-11-05 | 2023-10-24 | Corning Incorporated | Fixed bed bioreactor and methods of using the same |
| CN116814396A (en) * | 2023-06-29 | 2023-09-29 | 广州沙艾生物科技有限公司 | Stem cell culture device |
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