CN103614298A - Cell culture method and cell culture plate applied thereto - Google Patents
Cell culture method and cell culture plate applied thereto Download PDFInfo
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- CN103614298A CN103614298A CN201310667734.0A CN201310667734A CN103614298A CN 103614298 A CN103614298 A CN 103614298A CN 201310667734 A CN201310667734 A CN 201310667734A CN 103614298 A CN103614298 A CN 103614298A
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- culture plate
- tissue culture
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- 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
- C12M23/12—Well or multiwell plates
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- 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
- C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
- C12M27/02—Stirrer or mobile mixing elements
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Abstract
The invention provides a cell culture method and a cell culture plate applied thereto. The cell culture plate is characterized in that a stirring shaft is perpendicularly fixed in each deep culture hole of the cell culture plate; blades are fixed to each stirring shaft; the height of each blade is smaller than that of each hole; under the action of a certain external rotating speed, the cell culture plate and the blades synchronously move; the blades rotate relative to a culture solution and are used for stirring and mixing the culture solution in the cell culture plate. The volume dissolved oxygen coefficient of the cell culture plate can be increased and is approximate to that of a fermentation tank.
Description
Technical field
The present invention relates to cell culture apparatus technical field, relate in particular to the Tissue Culture Plate of a kind of cell culture processes and use thereof.
Background technology
The parallel fermentation technique of high-throughput is a kind of modern technique of studying fermenting process in corpusculum is culture apparatus, gropes fermentation control parameter, screening dominant strain, its result of study directly volume is amplified to technical scale, greatly reduce R&D costs, improve efficiency of research and development.
The parallel fermentation technique of current high-throughput mainly adopts Tissue Culture Plate or small-sized fermentation tank to carry out cell cultures.Tissue Culture Plate simple in structure, easy to operate, be widely used in bacterial screening, in use generally culture plate is placed on vibrator, by vibrator, provide certain rotating speed and eccentricity effect, make the nutrient solution vibration in culture plate, mix, but it is little that the gas-liquid of Tissue Culture Plate is practised physiognomy, liquid mixing effect is poor, volume oxygen transfer coefficient is significantly less than the volume oxygen transfer coefficient of fermentor tank, cause scale based on culture plate amplify can not Accurate Prediction fermentor tank in the cultivation results of aerobic cell, use the bacterial classification that culture plate filters out might not be applicable to fermentor tank simultaneously, simultaneously, each small-sized fermentation tank is equipped with motor, thereby drive the blade that is fixed on output terminal to rotate stir culture liquid by motor, the volume oxygen transfer coefficient of small-sized fermentation tank comparatively approaches the volume oxygen transfer coefficient of fermentor tank, but the complex structure of small-sized fermentation tank, manufacturing cost is high, and its use is limited by very large.
Summary of the invention
For the problems referred to above, the invention provides the Tissue Culture Plate of a kind of cell culture processes and use thereof, the volume dissolved oxygen coefficient that it can improve culture plate, reaches the volume dissolved oxygen coefficient level of approximation with fermentor tank.
Its technical scheme is such, a kind of cell culture processes, its culture apparatus is Tissue Culture Plate, it is characterized in that: each of described Tissue Culture Plate cultivated in deep hole and be vertically fixed with stir shaft, on described stir shaft, is fixed with blade, and the height of described blade is less than the height in hole, the external certain rotating speed providing act under, described Tissue Culture Plate and described blade are synchronized with the movement, and blade rotates with respect to nutrient solution, and the nutrient solution in Tissue Culture Plate is stirred, mixed.
It is further characterized in that:
The external certain rotating speed providing and eccentricity act under, described Tissue Culture Plate and described blade are synchronized with the movement, blade rotates with respect to nutrient solution, and the nutrient solution in Tissue Culture Plate is stirred, mixed.
The Tissue Culture Plate that above-mentioned cell culture processes is used, this Tissue Culture Plate is evenly distributed with cultivation deep hole, it is characterized in that: on the bottom face of described cultivation deep hole, be vertically installed with stir shaft, described stir shaft top is provided with blade, the height of described blade is less than the height in hole, described blade quantity is N, and N is random natural number and N >=1.
It is further characterized in that:
Described cultivation deep hole interior week baffle plate is installed, described baffle plate is at least two and uniform along the side face of described cultivation deep hole.
It is further characterised in that:
Described blade is contour layout in the hole depth direction of described cultivation deep hole;
Adjacent described blade is arranged in the hole depth direction dislocation of described cultivation deep hole;
Described baffle plate is contour layout in the hole depth direction of described cultivation deep hole;
Described blade quantity is 2N the outer radial periphery that is distributed in described stir shaft, adjacent two described blades layout that misplaces in the hole depth direction of described cultivation deep hole, two relative described blades contour being arranged in parallel in the hole depth direction of described cultivation deep hole;
Described blade is provided with four and be distributed in the outer radial periphery of described stir shaft, adjacent two described blades layout that misplaces in the hole depth direction of described cultivation deep hole, relative two described blades contour being arranged in parallel in the hole depth direction of described cultivation deep hole, described baffle plate is provided with four, four described baffle plates contour layout in the hole depth direction of described cultivation deep hole.
Adopt after the present invention, Tissue Culture Plate can be fixed on vibrator, vibrator provides certain rotating speed and eccentricity effect, cultivate in deep hole and be fixed with stir shaft, on stir shaft, be fixed with blade, Tissue Culture Plate and blade are synchronized with the movement, blade rotates with respect to nutrient solution, the effect that nutrient solution is played stirring, mixed, is dissolved in nutrient solution oxygen, has improved the volume dissolved oxygen coefficient of Tissue Culture Plate, simultaneously, Tissue Culture Plate of the present invention is avoided cultivating deep hole configuration motor into each, and it is simple in structure, greatly reduces its manufacturing cost; In the cultivation deep hole of Tissue Culture Plate, arrange after baffle plate, prevent that liquid level central authorities from producing eddy current, impel liquid fierceness to stir, improved the mixed effect of air and nutrient solution, further improved the volume dissolved oxygen coefficient of deep hole Tissue Culture Plate; The adjacent blades layout that misplaces in the hole depth direction of described cultivation deep hole, causes the nutrient solution adjacent blade of flowing through can change direction, has strengthened the mixing effect of nutrient solution, has further improved the volume dissolved oxygen coefficient of deep hole Tissue Culture Plate; In sum, Tissue Culture Plate of the present invention can improve the volume dissolved oxygen coefficient of culture plate, reaches the volume dissolved oxygen coefficient level of approximation with fermentor tank.
Accompanying drawing explanation
Fig. 1 is the plan structure schematic diagram of one embodiment of the invention;
Fig. 2 is the assembly structure schematic diagram of blade and stir shaft in Fig. 1;
Fig. 3 is the plan structure schematic diagram after single cultivation deep hole amplification in Fig. 1;
In Fig. 3 of Fig. 4, A-A is to structural representation;
Fig. 5 be in Fig. 3 B-B to structural representation;
Fig. 6 is the assembly structure schematic diagram of blade and stir shaft in another embodiment of the present invention.
Embodiment
A kind of cell culture processes, its culture apparatus is Tissue Culture Plate, Tissue Culture Plate is fixed on vibrator, each of Tissue Culture Plate cultivated in deep hole and is vertically fixed with stir shaft, on stir shaft, be fixed with blade, act on down to the certain rotating speed being provided and eccentricity by vibrator, Tissue Culture Plate and blade are synchronized with the movement, blade rotates with respect to nutrient solution, and the nutrient solution in Tissue Culture Plate is stirred, mixed.
As shown in Figures 1 to 5, this Tissue Culture Plate is evenly distributed with 24 and cultivates deep hole, be positioned at the axis direction of described cultivation deep hole, on the bottom face of described cultivation deep hole, be fixed with stir shaft, described stir shaft top is circumferentially evenly fixed with four blades, adjacent two described blades layout that misplaces in the hole depth direction of described cultivation deep hole, two relative described blades contour being arranged in parallel in the hole depth direction of described cultivation deep hole; Described cultivation deep hole be fixed with four baffle plates interior week, described baffle plate along the side face of described cultivation deep hole uniform and in the hole depth direction of described cultivation deep hole contour layout.
Substantially the same manner as Example 2, its difference is: described blade is contour layout in the hole depth direction of described cultivation deep hole.
As described in Figure 6, this Tissue Culture Plate is evenly distributed with 24 cultivation deep holes, be positioned at the axis direction of described cultivation deep hole, on the bottom face of described cultivation deep hole, be fixed with stir shaft, described stir shaft top is circumferentially evenly fixed with four blades, four blades contour layout in the hole depth direction of described cultivation deep hole; Described cultivation deep hole be fixed with four baffle plates interior week, described baffle plate along the side face of described cultivation deep hole uniform and in the hole depth direction of described cultivation deep hole contour layout.
Technique scheme has only embodied the preferred version of technical solution of the present invention, and those skilled in the art have all embodied principle of the present invention to some changes that wherein some part may be made, within belonging to the scope of protection of the invention.
Claims (9)
1. a cell culture processes, its culture apparatus is Tissue Culture Plate, it is characterized in that: each of described Tissue Culture Plate cultivated in deep hole and be vertically fixed with stir shaft, on described stir shaft, be fixed with blade, the height of described blade is less than the height in hole, the external certain rotating speed providing act under, described Tissue Culture Plate and described blade are synchronized with the movement, blade rotates with respect to nutrient solution, and the nutrient solution in Tissue Culture Plate is stirred, mixed.
2. a kind of cell culture processes according to claim 1, it is characterized in that: the external certain rotating speed providing and eccentricity act under, described Tissue Culture Plate and described blade are synchronized with the movement, blade rotates with respect to nutrient solution, and the nutrient solution in Tissue Culture Plate is stirred, mixed.
3. the Tissue Culture Plate that a kind of cell culture processes claimed in claim 1 is used, described Tissue Culture Plate is evenly distributed with cultivation deep hole, it is characterized in that: on the bottom face of described cultivation deep hole, be vertically installed with stir shaft, described stir shaft top is provided with blade, the height of described blade is less than the height in hole, described blade quantity is N, and N is random natural number and N >=1.
4. Tissue Culture Plate according to claim 3, is characterized in that: described cultivation deep hole interior week baffle plate is installed, described baffle plate is at least two and uniform along the side face of described cultivation deep hole.
5. according to the Tissue Culture Plate described in claim 3 or 4, it is characterized in that: described blade is contour layout in the hole depth direction of described cultivation deep hole.
6. according to the Tissue Culture Plate described in claim 3 or 4, it is characterized in that: adjacent described blade is arranged in the hole depth direction dislocation of described cultivation deep hole.
7. Tissue Culture Plate according to claim 4, is characterized in that: described baffle plate is contour layout in the hole depth direction of described cultivation deep hole.
8. according to the Tissue Culture Plate described in claim 3 or 4, it is characterized in that: described blade quantity is 2N the outer radial periphery that is distributed in described stir shaft, adjacent two described blades layout that misplaces in the hole depth direction of described cultivation deep hole, two relative described blades contour being arranged in parallel in the hole depth direction of described cultivation deep hole.
9. Tissue Culture Plate according to claim 4, it is characterized in that: described blade is provided with four and be distributed in the outer radial periphery of described stir shaft, adjacent two described blades layout that misplaces in the hole depth direction of described cultivation deep hole, relative two described blades contour being arranged in parallel in the hole depth direction of described cultivation deep hole, described baffle plate is provided with four, four described baffle plates contour layout in the hole depth direction of described cultivation deep hole.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201310667734.0A CN103614298B (en) | 2013-12-11 | 2013-12-11 | Cell culture method and cell culture plate applied thereto |
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| CN201310667734.0A CN103614298B (en) | 2013-12-11 | 2013-12-11 | Cell culture method and cell culture plate applied thereto |
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| CN103614298A true CN103614298A (en) | 2014-03-05 |
| CN103614298B CN103614298B (en) | 2015-04-29 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109415679A (en) * | 2016-06-30 | 2019-03-01 | 富士胶片株式会社 | The membrane separating method and cell culture apparatus of cell suspension |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1124292A (en) * | 1995-11-07 | 1996-06-12 | 许晓增 | Biological reactor for cell culture |
| JP2001231542A (en) * | 2000-02-22 | 2001-08-28 | Hideo Tanaka | Container for shaking culture and method for shaking culture |
| EP1884561A1 (en) * | 2005-03-22 | 2008-02-06 | Suntory Limited | Ventilating/stirring culture tank |
| WO2010125006A1 (en) * | 2009-04-28 | 2010-11-04 | Ge Healthcare Uk Limited | Method and apparatus for maintaining microcarrier beads in suspension |
| CN201648375U (en) * | 2009-12-29 | 2010-11-24 | 上海日泰医药设备工程有限公司 | Biological fermentation tank |
| CN203212577U (en) * | 2012-12-28 | 2013-09-25 | 江南大学 | Deep-hole cell culture plate with baffle plates |
-
2013
- 2013-12-11 CN CN201310667734.0A patent/CN103614298B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1124292A (en) * | 1995-11-07 | 1996-06-12 | 许晓增 | Biological reactor for cell culture |
| JP2001231542A (en) * | 2000-02-22 | 2001-08-28 | Hideo Tanaka | Container for shaking culture and method for shaking culture |
| EP1884561A1 (en) * | 2005-03-22 | 2008-02-06 | Suntory Limited | Ventilating/stirring culture tank |
| WO2010125006A1 (en) * | 2009-04-28 | 2010-11-04 | Ge Healthcare Uk Limited | Method and apparatus for maintaining microcarrier beads in suspension |
| CN201648375U (en) * | 2009-12-29 | 2010-11-24 | 上海日泰医药设备工程有限公司 | Biological fermentation tank |
| CN203212577U (en) * | 2012-12-28 | 2013-09-25 | 江南大学 | Deep-hole cell culture plate with baffle plates |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109415679A (en) * | 2016-06-30 | 2019-03-01 | 富士胶片株式会社 | The membrane separating method and cell culture apparatus of cell suspension |
| US11492578B2 (en) | 2016-06-30 | 2022-11-08 | Fujifilm Corporation | Membrane separation method of cell suspension, and cell culture device |
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| CN103614298B (en) | 2015-04-29 |
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Effective date of registration: 20210826 Address after: 610000 South Sixth Road 669, Chengdu Economic and Technological Development Zone, Sichuan Province Patentee after: CHENGDU YINGDE BIO-MEDICAL EQUIPMENT TECHNOLOGY Co.,Ltd. Patentee after: Jiangnan University Address before: 213000 d3010, No.26 Huashan Middle Road, Xinbei District, Changzhou City, Jiangsu Province Patentee before: CHANGZHOU YINGDE BIOTECHNOLOGY Co.,Ltd. Patentee before: Jiangnan University |
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