CN113337400A - Stem cell culture bottle support device and using method - Google Patents
Stem cell culture bottle support device and using method Download PDFInfo
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- CN113337400A CN113337400A CN202110596068.0A CN202110596068A CN113337400A CN 113337400 A CN113337400 A CN 113337400A CN 202110596068 A CN202110596068 A CN 202110596068A CN 113337400 A CN113337400 A CN 113337400A
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- slider
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- 238000000034 method Methods 0.000 title claims abstract description 11
- 210000000130 stem cell Anatomy 0.000 title claims abstract description 10
- 238000004113 cell culture Methods 0.000 title claims abstract description 8
- 229910000838 Al alloy Inorganic materials 0.000 claims 1
- 239000000463 material Substances 0.000 claims 1
- 239000010935 stainless steel Substances 0.000 claims 1
- 229910001220 stainless steel Inorganic materials 0.000 claims 1
- 210000004027 cell Anatomy 0.000 abstract description 14
- 239000007788 liquid Substances 0.000 abstract description 5
- 238000003860 storage Methods 0.000 abstract description 3
- 239000002699 waste material Substances 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 7
- 238000004140 cleaning Methods 0.000 description 5
- 102000038379 digestive enzymes Human genes 0.000 description 4
- 108091007734 digestive enzymes Proteins 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 210000002901 mesenchymal stem cell Anatomy 0.000 description 3
- 230000029087 digestion Effects 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000004114 suspension culture Methods 0.000 description 1
- 210000003954 umbilical cord Anatomy 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
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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
- C12M23/48—Holding appliances; Racks; Supports
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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
- C12M23/08—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
- C12M27/00—Means for mixing, agitating or circulating fluids in the vessel
- C12M27/16—Vibrating; Shaking; Tilting
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- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Sustainable Development (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
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- Clinical Laboratory Science (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
本发明涉及干细胞领域，具体提供一种干细胞培养瓶支架装置及使用方法。该支架装置由侧支架和活动支架组成，侧支架为固定不动架子，起到整体支撑作用。活动支架设有固定挡板和滑块组成的可调节大小的培养瓶存放区，可沿一侧连在侧支架上的轴顺时针和逆时针旋转10度，起到摇晃混匀培养瓶和使其中的液体集中在底部的作用；活动支架分三层，每层可放四个培养瓶。使用时将培养瓶置于活动支架的培养瓶放置区，然后整体置于生物安全柜和超净工作台中，利用活动支架的可旋转性完成废液的移除、加液的混匀、细胞的收集等。本培养瓶支架系统可有效辅助完成批量培养瓶换液、细胞传代和收集操作，使整个过程便利化，节约大量时间。The invention relates to the field of stem cells, and in particular provides a stem cell culture flask support device and a method for using the same. The bracket device is composed of a side bracket and a movable bracket, and the side bracket is a fixed and immovable shelf, which plays an integral supporting role. The movable bracket is provided with an adjustable-size culture bottle storage area composed of a fixed baffle plate and a slider, which can be rotated 10 degrees clockwise and counterclockwise along the shaft connected to the side bracket on one side to shake and mix the culture bottle and make the bottle evenly. The liquid in it is concentrated at the bottom; the movable bracket is divided into three layers, and each layer can hold four culture flasks. When in use, place the culture bottle in the culture bottle placement area of the movable stand, and then place it in the biological safety cabinet and ultra-clean workbench as a whole, and use the rotatability of the movable stand to complete the removal of waste liquid, the mixing of adding liquid, and the removal of cells. collection etc. The culture flask support system can effectively assist in the completion of batch culture flask exchange, cell passage and collection operations, which facilitates the entire process and saves a lot of time.
Description
Field of the invention
The invention belongs to the field of stem cells, and particularly relates to a stem cell culture bottle support device and a using method thereof.
Background
With the development of mesenchymal stem cell storage and bank establishment and clinical application research, batch expansion has become a main mode of stem cell production. Currently, mesenchymal stem cells are expanded in batches in a plurality of optional modes, such as cell factories, 3D micro-slide suspension culture and the like, but culture flask culture is still the most important mode at present due to the advantages of simple operation, high safety and the like. The number of flasks varies from tens to tens of flasks per operation. Under conventional conditions, each flask is subjected to the following operations during the passage or collection of cells: the method comprises the steps of taking up a bottle, pouring out a culture medium, adding a cleaning solution for cleaning, pouring out the cleaning solution, adding pancreatin for digestion (shaking up), putting down the bottle, blowing up cells and transferring the cells to a centrifuge tube, and the like.
Disclosure of Invention
The invention provides a cell culture bottle support device and a use method thereof, the device consists of a side support (4) and a movable support (5), and 12 culture bottles can be placed at the same time for operation. The device can realize batch operation to the blake bottle of different specifications: the operations of removing supernatant, cleaning cells, adding enzyme, mixing and the like are convenient and quick in the whole process, and a large amount of time is saved.
The side brackets (4) are fixed brackets and are positioned at two ends of the culture bottle bracket; each of the movable supports is composed of a rectangular base and a rectangular vertical plate, three groups of holes are arranged on the vertical plate in parallel to the bottom plate, each group has two holes, one hole is circular and is used for connecting a rear shaft of the movable support, fixing the movable support and providing a rotary support for the movable support; the other hole is an arc-shaped hole and is a connecting position of a rotating shaft screw rod (12) of the movable support, and the rotating shaft screw rod of the movable support can move up and down along an arc line to adjust the inclined position of the movable support. The whole side bracket plays a role in fixing the culture bottle bracket.
Above-mentioned movable support (5) comprise three slotted litter, are equipped with fixed stop (1) and slider (2) on the litter, and every fixed stop forms an adjustable blake bottle storage position with corresponding the slider, and fixed stop can not remove, and the slider can move on the spout, thereby adjusts the distance between slider and the fixed stop and can be fit for placing not equidimension blake bottle, and four blake bottles can be put to every movable support.
The movable support is provided with a shaft along each of two sides of the long shaft, two ends of the rear shaft are placed on the fixed support, the front shaft is a movable support rotating shaft screw rod and can move in the arc-shaped groove on the fixed support, so that the whole movable support rotates clockwise and anticlockwise by taking the rear shaft as a center, and the inclined position of the movable support is adjusted.
The movable support has three groups, each group can be used for placing 4 culture bottles, and the three groups can be used for placing 12 culture bottles.
The using method comprises the following steps:
1) the position of the movable block is adjusted to ensure that the size between the sliding block and the fixed baffle is suitable for the size of the culture bottle.
2) The culture flask is taken out of the incubator, placed on a rack and transferred to a biological safety cabinet or an ultra-clean workbench.
3) Opening the culture bottle, moving the movable bracket rotating shaft screw rod anticlockwise to enable the culture product to incline at a certain angle, collecting the culture solution at the bottom of the bottle, and removing all the culture medium by using a pipette.
4) Adding physiological saline or PBS, moving the movable support rotating shaft screw clockwise and anticlockwise for 2-3 times to clean cells, moving the movable support rotating shaft screw anticlockwise to enable the culture bottle to incline at a certain angle, collecting culture solution at the bottom of the bottle, and removing all culture medium by using a pipette.
5) Adding digestive enzyme, moving the movable support rotating shaft screw clockwise and anticlockwise for 2-3 times to uniformly distribute the digestive enzyme in the whole culture bottle, stopping at the horizontal position of the movable support, and incubating and digesting for 3-5 minutes at room temperature.
6) Adding serum, moving the movable support rotating shaft screw clockwise and anticlockwise for 2-3 times to uniformly distribute the serum in the whole culture bottle to stop digestion, and moving the movable support rotating shaft screw anticlockwise to enable the culture bottle to incline at a certain angle and collect liquid at the bottom of the bottle.
7) The cells are detached from the bottle wall by pipetting and dispersed into the liquid, and the cell suspension is aspirated.
Drawings
FIG. 1 front view of a culture flask holder
FIG. 2 side view of a culture flask holder
FIG. 3 three-dimensional drawing of a culture flask holder
FIG. 4 is a diagram showing the three-dimensional effect of the inclined movable support
Examples
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.
Example 1 umbilical cord mesenchymal stem cells were cultured in T75 culture flasks, 20 flasks in total, and the cells were subcultured with the digested cells. Conventionally, each flask of cells requires the following steps (the cap has been unscrewed): picking up the culture flask (5 seconds), pouring off the culture medium (5 seconds), adding a cleaning solution for washing (10 seconds), adding a digestive enzyme (10 seconds), and putting down the culture flask (5 seconds), wherein 35 seconds are required for 20 flasks of cells, and 700 seconds are required for 20 flasks of cells; the culture bottle bracket operation is adopted (the bottle cap is unscrewed, 10 bottles are operated at a time): the medium was aspirated off (4 sec/vial for 40 sec), the wash solution was added (4 sec/vial for 40 sec), the wash solution was aspirated off (4 sec/vial for 40 sec), and the digestive enzymes were added (4 sec/vial for 40 sec), for 160 sec and for 320 sec for 20 vials. Compared with the conventional operation, the culture bottle bracket operation can save half time.
Claims (11)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110596068.0A CN113337400A (en) | 2021-05-31 | 2021-05-31 | Stem cell culture bottle support device and using method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110596068.0A CN113337400A (en) | 2021-05-31 | 2021-05-31 | Stem cell culture bottle support device and using method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113337400A true CN113337400A (en) | 2021-09-03 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110596068.0A Pending CN113337400A (en) | 2021-05-31 | 2021-05-31 | Stem cell culture bottle support device and using method |
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| CN (1) | CN113337400A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2025130872A1 (en) * | 2023-12-18 | 2025-06-26 | 浙江鲁墨科学仪器有限公司 | Culture device, culture vessel, and lid assembly |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN207608547U (en) * | 2017-12-11 | 2018-07-13 | 齐齐哈尔医学院 | Cell culture culture bottle fixed frame |
| CN208933325U (en) * | 2018-05-19 | 2019-06-04 | 管乐敏 | A kind of dedicated placement bracket of Tissue Culture Flask |
| CN209481642U (en) * | 2018-09-06 | 2019-10-11 | 科美(南京)智能科技有限公司 | A kind of Tissue Culture Flask automatic adjustment platform |
| CN210560359U (en) * | 2019-03-11 | 2020-05-19 | 河北森朗生物科技有限公司 | Cell culture bottle storage rack |
| CN210596095U (en) * | 2019-08-13 | 2020-05-22 | 云南新生命干细胞有限公司 | Cell culture bottle rack |
| WO2021026272A1 (en) * | 2019-08-07 | 2021-02-11 | Becton, Dickinson And Company | High density bottle drum for storage, agitation and reading of blood culture bottles and methods of storing |
| CN212855839U (en) * | 2020-07-20 | 2021-04-02 | 河南中科干细胞基因工程有限公司 | Cell culture bottle storage rack |
| CN215517473U (en) * | 2021-08-27 | 2022-01-14 | 北京艾颐生物科技有限公司 | Stem cell culture perfusion device |
-
2021
- 2021-05-31 CN CN202110596068.0A patent/CN113337400A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN207608547U (en) * | 2017-12-11 | 2018-07-13 | 齐齐哈尔医学院 | Cell culture culture bottle fixed frame |
| CN208933325U (en) * | 2018-05-19 | 2019-06-04 | 管乐敏 | A kind of dedicated placement bracket of Tissue Culture Flask |
| CN209481642U (en) * | 2018-09-06 | 2019-10-11 | 科美(南京)智能科技有限公司 | A kind of Tissue Culture Flask automatic adjustment platform |
| CN210560359U (en) * | 2019-03-11 | 2020-05-19 | 河北森朗生物科技有限公司 | Cell culture bottle storage rack |
| WO2021026272A1 (en) * | 2019-08-07 | 2021-02-11 | Becton, Dickinson And Company | High density bottle drum for storage, agitation and reading of blood culture bottles and methods of storing |
| CN210596095U (en) * | 2019-08-13 | 2020-05-22 | 云南新生命干细胞有限公司 | Cell culture bottle rack |
| CN212855839U (en) * | 2020-07-20 | 2021-04-02 | 河南中科干细胞基因工程有限公司 | Cell culture bottle storage rack |
| CN215517473U (en) * | 2021-08-27 | 2022-01-14 | 北京艾颐生物科技有限公司 | Stem cell culture perfusion device |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2025130872A1 (en) * | 2023-12-18 | 2025-06-26 | 浙江鲁墨科学仪器有限公司 | Culture device, culture vessel, and lid assembly |
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Application publication date: 20210903 |
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