CN103614298B - 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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- CN103614298B CN103614298B CN201310667734.0A CN201310667734A CN103614298B CN 103614298 B CN103614298 B CN 103614298B CN 201310667734 A CN201310667734 A CN 201310667734A CN 103614298 B CN103614298 B CN 103614298B
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- culture plate
- blade
- deep hole
- cell 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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- Chemical & Material Sciences (AREA)
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- Biomedical Technology (AREA)
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- Apparatus Associated With Microorganisms And Enzymes (AREA)
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, particularly relate 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 studied fermenting process in little system culture apparatus, grope ferment control parameter, screen dominant strain, its result of study directly can be amplified to technical scale by volume, 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.The structure of Tissue Culture Plate is simple, easy to operate, be widely used in bacterial screening, in use general culture plate is placed on vibrator, certain rotating speed and eccentricity effect is provided by vibrator, nutrient solution in culture plate is vibrated, mixing, but the gas-liquid of Tissue Culture Plate is practised physiognomy little, liquid mixing effect is poor, volume oxygen transfer coefficient is significantly less than the volume oxygen transfer coefficient of fermentor tank, causing the scale based on culture plate to be amplified can not the cultivation results of aerobic cell in Accurate Prediction fermentor tank, the bacterial classification simultaneously using culture plate to filter out might not be applicable to fermentor tank, simultaneously, each small-sized fermentation tank is equipped with motor, the blade being fixed on output terminal is driven to rotate thus stir culture liquid by motor, the volume oxygen transfer coefficient of small-sized fermentation tank is comparatively close to 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, it can improve the volume dissolved oxygen coefficient of 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: be vertically fixed with stir shaft in each cultivation deep hole of described Tissue Culture Plate, described stir shaft is fixed with blade, the height of described blade is less than the height in hole, the external certain rotating speed provided act under, described Tissue Culture Plate and described blade are synchronized with the movement, and blade rotates relative to nutrient solution, stir the nutrient solution in Tissue Culture Plate, mix.
It is further characterized in that:
External certain rotating speed of providing and eccentricity act under, described Tissue Culture Plate and described blade are synchronized with the movement, and blade rotates relative to nutrient solution, stir the nutrient solution in Tissue Culture Plate, mix.
The Tissue Culture Plate that above-mentioned cell culture processes uses, this Tissue Culture Plate is evenly distributed with cultivation deep hole, it is characterized in that: the bottom face of described cultivation deep hole is 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 that N, N are random natural number and N >=1.
It is further characterized in that:
The inner circumferential of described cultivation deep hole is provided with baffle plate, and described baffle plate is at least two pieces and side face along described cultivation deep hole is uniform.
It is further characterised in that:
Described blade contour layout on the hole depth direction of described cultivation deep hole;
Adjacent described blade is in the hole depth direction of described cultivation deep hole dislocation layout;
Described baffle plate contour layout on the hole depth direction of described cultivation deep hole;
Described blade quantity is 2N and is distributed in the outer radial periphery of described stir shaft, and blade described in adjacent two panels misplaces layout on the hole depth direction of described cultivation deep hole, and blade described in relative two panels is contour on the hole depth direction of described cultivation deep hole to be arranged in parallel;
Described blade is provided with four and is distributed in the outer radial periphery of described stir shaft, blade described in adjacent two panels misplaces layout on the hole depth direction of described cultivation deep hole, blade described in relative two panels is contour on the hole depth direction of described cultivation deep hole to be arranged in parallel, described baffle plate is provided with four pieces, four pieces of described baffle plates contour layout on the hole depth direction of described cultivation deep hole.
After adopting the present invention, Tissue Culture Plate can be fixed on vibrator, vibrator provides certain rotating speed and eccentricity effect, cultivate deep hole internal fixtion and have stir shaft, stir shaft is fixed with blade, Tissue Culture Plate and blade are synchronized with the movement, blade rotates relative to nutrient solution, nutrient solution is played to the effect of stirring, mixing, make oxygen be dissolved in nutrient solution, improve the volume dissolved oxygen coefficient of Tissue Culture Plate, simultaneously, Tissue Culture Plate of the present invention is avoided as each cultivation deep hole configuration motor, and its structure is simple, greatly reduces its manufacturing cost; After baffle plate is set in the cultivation deep hole of Tissue Culture Plate, prevents liquid level central authorities from producing eddy current, impel liquid fierceness to stir, improve the mixed effect of air and nutrient solution, further increase the volume dissolved oxygen coefficient of Deep-hole cell culture plate; Adjacent blades misplaces layout on the hole depth direction of described cultivation deep hole, and causing nutrient solution to flow through adjacent blade can change direction, strengthens the mixing effect of nutrient solution, further improves the volume dissolved oxygen coefficient of Deep-hole cell 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 in Fig. 1 after single cultivation deep hole amplification;
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
Embodiment 1
A kind of cell culture processes, its culture apparatus is Tissue Culture Plate, Tissue Culture Plate is fixed on vibrator, vertically stir shaft is fixed with in each cultivation deep hole of Tissue Culture Plate, stir shaft is fixed with blade, and act on down to the certain rotating speed provided by vibrator and eccentricity, Tissue Culture Plate and blade are synchronized with the movement, blade rotates relative to nutrient solution, stirs the nutrient solution in Tissue Culture Plate, mixes.
Embodiment 2
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, the bottom face of described cultivation deep hole is fixed with stir shaft, described stir shaft top circumference is evenly fixed with four blades, blade described in adjacent two panels misplaces layout on the hole depth direction of described cultivation deep hole, and blade described in relative two panels is contour on the hole depth direction of described cultivation deep hole to be arranged in parallel; The inner circumferential of described cultivation deep hole is fixed with four pieces of baffle plates, described baffle plate along the side face of described cultivation deep hole uniform and on the hole depth direction of described cultivation deep hole contour layout.
Embodiment 3
Substantially the same manner as Example 2, its difference is: described blade contour layout on 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, the bottom face of described cultivation deep hole is fixed with stir shaft, described stir shaft top circumference is evenly fixed with four blades, four blades contour layout on the hole depth direction of described cultivation deep hole; The inner circumferential of described cultivation deep hole is fixed with four pieces of baffle plates, described baffle plate along the side face of described cultivation deep hole uniform and on the hole depth direction of described cultivation deep hole contour layout.
Technique scheme only embodies the preferred version of technical solution of the present invention, and those skilled in the art all embody principle of the present invention to some variations that wherein some part may be made, and belong within the scope of protection of the invention.
Claims (2)
1. a Tissue Culture Plate, described Tissue Culture Plate is evenly distributed with cultivation deep hole, it is characterized in that: the bottom face of described cultivation deep hole is vertically installed with stir shaft, and described stir shaft top is provided with blade, and the height of described blade is less than the height in hole; The inner circumferential of described cultivation deep hole is provided with baffle plate, and described baffle plate is at least two pieces and side face along described cultivation deep hole is uniform; Described blade is provided with four and is distributed in the outer radial periphery of described stir shaft, blade described in adjacent two panels misplaces layout on the hole depth direction of described cultivation deep hole, blade described in relative two panels is contour on the hole depth direction of described cultivation deep hole to be arranged in parallel, described baffle plate is provided with four pieces, four pieces of described baffle plates contour layout on the hole depth direction of described cultivation deep hole.
2. a cell culture processes, it is characterized in that: utilize the Tissue Culture Plate described in claim 1, under acting on to the certain rotating speed provided at vibrator and eccentricity, described Tissue Culture Plate and described blade are synchronized with the movement, blade rotates relative to nutrient solution, stirs the nutrient solution in Tissue Culture Plate, mixes.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310667734.0A CN103614298B (en) | 2013-12-11 | 2013-12-11 | Cell culture method and cell culture plate applied thereto |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310667734.0A CN103614298B (en) | 2013-12-11 | 2013-12-11 | Cell culture method and cell culture plate applied thereto |
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| Publication Number | Publication Date |
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| CN103614298A CN103614298A (en) | 2014-03-05 |
| CN103614298B true CN103614298B (en) | 2015-04-29 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201310667734.0A Active CN103614298B (en) | 2013-12-11 | 2013-12-11 | Cell culture method and cell culture plate applied thereto |
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Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2018003476A1 (en) | 2016-06-30 | 2018-01-04 | 富士フイルム株式会社 | Cell-suspension membrane separation method and cell culture device |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1124292A (en) * | 1995-11-07 | 1996-06-12 | 许晓增 | Biological reactor for cell culture |
| EP1884561A1 (en) * | 2005-03-22 | 2008-02-06 | Suntory Limited | Ventilating/stirring culture tank |
| 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 |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001231542A (en) * | 2000-02-22 | 2001-08-28 | Hideo Tanaka | Container for shaking culture and method for shaking culture |
| GB0907260D0 (en) * | 2009-04-28 | 2009-06-10 | Ge Healthcare Uk Ltd | Method and apparatus for maintaining microcarrier beads in suspension |
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2013
- 2013-12-11 CN CN201310667734.0A patent/CN103614298B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1124292A (en) * | 1995-11-07 | 1996-06-12 | 许晓增 | Biological reactor for cell culture |
| EP1884561A1 (en) * | 2005-03-22 | 2008-02-06 | Suntory Limited | Ventilating/stirring culture tank |
| 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 |
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| CN103614298A (en) | 2014-03-05 |
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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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