CN110079459A - The adjusting method of gas balance in a kind of stem cell culture vessel - Google Patents
The adjusting method of gas balance in a kind of stem cell culture vessel Download PDFInfo
- Publication number
- CN110079459A CN110079459A CN201910403665.XA CN201910403665A CN110079459A CN 110079459 A CN110079459 A CN 110079459A CN 201910403665 A CN201910403665 A CN 201910403665A CN 110079459 A CN110079459 A CN 110079459A
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- Prior art keywords
- stem cell
- gas
- culture vessel
- cell culture
- carbon dioxide
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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
-
- 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
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/14—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus with filters, sieves or membranes
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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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/30—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration
- C12M41/34—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of gas
Abstract
The adjusting method of gas balance in a kind of stem cell culture vessel, the present invention relates to stem cell fields, and in particular to the adjusting method of gas balance in a kind of stem cell culture vessel.The problem of can not completing the quick discharge of oxygen being sufficiently fed with carbon dioxide according to the upgrowth situation of stem cell the present invention is to solve the culture of existing stem cell.Method: the sensor of carbon dioxide and oxygen is set in closed stem cell culture vessel, oxygen and carbon dioxide effectively are provided for stem cell according to specific requirements of the stem cell growth to carbon dioxide and oxygen, guarantee the gas balance of stem cell Fei Gennei, by regulating and controlling the gas balance of stem cell culture vessel, stem cell growth is effectively facilitated.Adjusting of the present invention for gas balance in stem cell culture vessel.
Description
Technical field
The present invention relates to stem cell fields, and in particular to the adjusting method of gas balance in a kind of stem cell culture vessel.
Background technique
In recent years, international, domestic stem cell market development is rapid, and stem cell clinical treatment is done by public recognition and acceptance
Cell requirements are constantly promoted.Stem cell cultivates 2D to 3D development, makes great efforts the output for expanding stem cell.It is cultivated in the 3D of stem cell
In system, in order to be effectively ensured stem cell sufficiently grow it is necessary that oxygen in cultivating system (stem cell culture vessel) with
The balance of carbon dioxide, it is necessary to the quick row of oxygen being sufficiently fed with carbon dioxide is completed according to the upgrowth situation of stem cell
Out.Current stem cell culture is all that the simple air by mixing carbon dioxide is completed, not conscientiously according to stem cell
Growth demand carries out gas supply.
Summary of the invention
The invention aims to solve existing stem cell culture not completing oxygen according to the upgrowth situation of stem cell
The problem of being sufficiently fed the quick discharge with carbon dioxide, and a kind of adjusting side of gas balance in stem cell culture vessel is provided
Method.
The adjusting method of gas balance specifically sequentially includes the following steps: in a kind of stem cell culture vessel of the present invention
One, stem cell to be cultivated is placed in stem cell culture vessel, oxygen sensor is set in stem cell culture vessel
Device and carbon dioxide sensor, while gas-exchange membrane is set in the culture solution inlet of stem cell culture vessel, then treat
Culture stem cell is cultivated, control nutrient solution circulation, and oxygen and carbon dioxide contains in real-time monitoring stem cell culture vessel
Amount；
Two, the oxygen partial pressure and carbon dioxide for regulating and controlling the gas side of gas-exchange membrane by air pump divide to change culture solution
The content of middle oxygen and carbon dioxide；
Three, the gas content in stem cell culture vessel is changed into partial pressure corresponding with the gas side of gas-exchange membrane
It is associated, when the consumption of stem cell oxygen increases, carbon dioxide forms enhancing, increase the content of oxygen in the culture solution of input,
Carbon dioxide content is reduced, guarantees the gas balance of stem cell growth environment.
Beneficial effects of the present invention:
The present invention effectively facilitates stem cell growth by the gas balance of regulation stem cell culture vessel, improves stem cell
Quality carries out stem-cell therapy by stem cell application specification input blood samples of patients.Stem cell is placed in one using the invention patent
Cultivated in a circulator, the actual consumption by monitoring stem cell growth process adjust in time oxygen content in culture solution with
Carbon dioxide content guarantees the gas balance of stem cell growth process, accelerates stem cell growth, can improve the growth speed of stem cell
Rate 5~10%.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of stem cell culture vessel.
Specific embodiment
Specific embodiment 1: the adjusting method of gas balance is specifically pressed in a kind of stem cell culture vessel of present embodiment
Following steps carry out:
One, stem cell to be cultivated is placed in stem cell culture vessel, oxygen sensor is set in stem cell culture vessel
Device and carbon dioxide sensor, while gas-exchange membrane is set in the culture solution inlet of stem cell culture vessel, then treat
Culture stem cell is cultivated, control nutrient solution circulation, and oxygen and carbon dioxide contains in real-time monitoring stem cell culture vessel
Amount；
Two, the oxygen partial pressure and carbon dioxide for regulating and controlling the gas side of gas-exchange membrane by air pump divide to change culture solution
The content of middle oxygen and carbon dioxide；
Three, the gas content in stem cell culture vessel is changed into partial pressure corresponding with the gas side of gas-exchange membrane
It is associated, when the consumption of stem cell oxygen increases, carbon dioxide forms enhancing, increase the content of oxygen in the culture solution of input,
Carbon dioxide content is reduced, guarantees the gas balance of stem cell growth environment.
Oxygen sensor described in present embodiment and carbon dioxide sensor be commercially available oxygen sensor O2-G2 and
MH-Z14, MH-Z18, MH-Z19 carbon dioxide sensor.
Specific embodiment 2: the present embodiment is different from the first embodiment in that: gas described in step 1 is handed over
Changing film is the air penetrating film that selectivity penetrates oxygen and carbon dioxide.Other are same as the specific embodiment one.
The selectivity is hollow-fibre membrane through the air penetrating film of oxygen and carbon dioxide, and gas and culture solution are in film
Two sides carry out the exchange of oxygen and carbon dioxide by diffusion, and this exchanged form avoids gas liquid and directly contacts.
Film used in present embodiment is provided by Wei Gao company.
Specific embodiment 3: the present embodiment is different from the first and the second embodiment in that: the stem cell culture
Container is tubulose, is made of tube body 2 and 1 two parts of lid, and tube body 2 and lid 1 pass through helixseal；It is set in the bottom of tube body 2
It is equipped with a culture solution entrance 4, has a culture solution outlet 3 in the center of lid 1, gas-exchange membrane 5 is rolled into tubular setting and exists
Culture solution entrance 4 is divided into gas side and hydraulic fluid side by the shaft core position of culture solution entrance 4, the gas-exchange membrane 5, and gas is handed over
The inside for changing film 5 is gas side, and the outside of gas-exchange membrane 5 is hydraulic fluid side.Other are the same as one or two specific embodiments.
Specific embodiment 4: unlike one of present embodiment and specific embodiment one to three: right in step 1
The content of initial carbon dioxide is controlled 10~15% when cultivating stem cell and being cultivated.Other with specific embodiment one to
One of three is identical.
Specific embodiment 5: unlike one of present embodiment and specific embodiment one to four: institute in step 2
Stating gas balance is to guarantee that carbon dioxide content is 10~15% in stem cell culture vessel by real-time monitoring.Other and tool
One of body embodiment one to four is identical.
Beneficial effects of the present invention are verified using following embodiment:
Embodiment one: the adjusting method of gas balance specifically sequentially includes the following steps: in a kind of stem cell culture vessel
One, NK cell is placed in stem cell culture vessel, oxygen sensor and two is set in stem cell culture vessel
Carbon sensor is aoxidized, while gas-exchange membrane is set in the culture solution inlet of stem cell culture vessel, it is dry then to treat culture
Cell is cultivated, control nutrient solution circulation, the content of oxygen and carbon dioxide in real-time monitoring stem cell culture vessel；Oxygen
Control is in 40mL/100mL；The content of initial carbon dioxide is controlled 10~15%；
Two, it is divided by the oxygen partial pressure and carbon dioxide that regulate and control the gas side of gas-exchange membrane to change oxygen in culture solution
The content of gas and carbon dioxide；
Three, the gas content in stem cell culture vessel is changed into partial pressure corresponding with the gas side of gas-exchange membrane
It is associated, when the consumption of stem cell oxygen increases, carbon dioxide forms enhancing, increase the content of oxygen in the culture solution of input,
Carbon dioxide content is reduced, guarantees the gas balance of stem cell growth environment.
The present embodiment effectively facilitates stem cell growth by the gas balance of regulation stem cell culture vessel, improves dry thin
Cytoplasm amount carries out stem-cell therapy by stem cell application specification input blood samples of patients.Stem cell is placed in using the invention patent
It is cultivated in one circulator, the actual consumption by monitoring stem cell growth process adjusts the oxygen content in culture solution in time
With carbon dioxide content, guarantee the gas balance of stem cell growth process, accelerates stem cell growth, the growth of stem cell can be improved
Rate 5~10%.
Claims (5)
1. the adjusting method of gas balance in a kind of stem cell culture vessel, it is characterised in that gas is flat in stem cell culture vessel
The adjusting method of weighing apparatus specifically sequentially includes the following steps:
One, stem cell to be cultivated is placed in stem cell culture vessel, in stem cell culture vessel be arranged oxygen sensor and
Carbon dioxide sensor, while gas-exchange membrane is set in the culture solution inlet of stem cell culture vessel, then treat culture
Stem cell is cultivated, control nutrient solution circulation, the content of oxygen and carbon dioxide in real-time monitoring stem cell culture vessel；
Two, the oxygen partial pressure and carbon dioxide for regulating and controlling the gas side of gas-exchange membrane by air pump divide to change oxygen in culture solution
The content of gas and carbon dioxide；
Three, the gas content variation partial pressure corresponding to the gas side of gas-exchange membrane in stem cell culture vessel is related
Connection increases the content of oxygen in the culture solution of input, reduces when the consumption of stem cell oxygen increases, carbon dioxide forms enhancing
Carbon dioxide content guarantees the gas balance of stem cell growth environment.
2. the adjusting method of gas balance in a kind of stem cell culture vessel according to claim 1, it is characterised in that step
Gas-exchange membrane described in rapid one is the air penetrating film that selectivity penetrates oxygen and carbon dioxide.
3. the adjusting method of gas balance in a kind of stem cell culture vessel according to claim 1, it is characterised in that institute
Stating stem cell culture vessel is tubulose, is made of tube body (2) and lid (1) two parts, and tube body (2) and lid (1) pass through spiral
Sealing；The bottom of tube body (2) is provided with a culture solution entrance (4), has a culture solution outlet in the center of lid (1)
(3), gas-exchange membrane (5) is rolled into tubular setting in the shaft core position of culture solution entrance (4), and the gas-exchange membrane (5) will train
Nutrient solution entrance (4) is divided into gas side and hydraulic fluid side, and the inside of gas-exchange membrane (5) is gas side, gas-exchange membrane (5) it is outer
Portion is hydraulic fluid side.
4. the adjusting method of gas balance in a kind of stem cell culture vessel according to claim 1, it is characterised in that step
The content that initial carbon dioxide when culture stem cell is cultivated is treated in rapid one is controlled 10~15%.
5. the adjusting method of gas balance in a kind of stem cell culture vessel according to claim 1, it is characterised in that step
Gas balance described in rapid two is to guarantee that carbon dioxide content is 10~15% in stem cell culture vessel by real-time monitoring.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113564125A (en) * | 2021-08-04 | 2021-10-29 | 河南省肿瘤医院 | Method for culturing tumor cells |
Citations (4)
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|---|---|---|---|---|
| US20100015660A1 (en) * | 2005-06-30 | 2010-01-21 | Synexa Life Sciences (Proprietary ) Limited | Production of Recombiant Products Using Capillary Membranes |
| WO2010040302A1 (en) * | 2008-10-10 | 2010-04-15 | 深圳市嘉天源生物科技有限公司 | Methods for isolating mesenchymal stem cells from embryos of human or animals and extracting secretion substances thereof |
| WO2011102531A1 (en) * | 2010-02-16 | 2011-08-25 | Kyoto University | Method of efficiently establishing induced pluripotent stem cells |
| CN103589808A (en) * | 2013-11-11 | 2014-02-19 | 中国人民解放军第三军医大学第一附属医院 | Automatic control method and system for pH/oxygen partial pressure/partial pressure of carbon dioxide of cell and tissue culture solution |
-
2019
- 2019-05-15 CN CN201910403665.XA patent/CN110079459A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100015660A1 (en) * | 2005-06-30 | 2010-01-21 | Synexa Life Sciences (Proprietary ) Limited | Production of Recombiant Products Using Capillary Membranes |
| WO2010040302A1 (en) * | 2008-10-10 | 2010-04-15 | 深圳市嘉天源生物科技有限公司 | Methods for isolating mesenchymal stem cells from embryos of human or animals and extracting secretion substances thereof |
| WO2011102531A1 (en) * | 2010-02-16 | 2011-08-25 | Kyoto University | Method of efficiently establishing induced pluripotent stem cells |
| CN103589808A (en) * | 2013-11-11 | 2014-02-19 | 中国人民解放军第三军医大学第一附属医院 | Automatic control method and system for pH/oxygen partial pressure/partial pressure of carbon dioxide of cell and tissue culture solution |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113564125A (en) * | 2021-08-04 | 2021-10-29 | 河南省肿瘤医院 | Method for culturing tumor cells |
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| TA01 | Transfer of patent application right |
Effective date of registration: 20200225 Address after: Room b1-5675, building 3, No.20 Yong'an Road, Shilong Economic Development Zone, Mentougou District, Beijing 102300 Applicant after: BEIJING HUANIU SHIJI BIOTECHNOLOGY RESEARCH INSTITUTE Address before: 116600 Hongmei District 32, Dalian Economic and Technological Development Zone, Liaoning Province, 1-4-2 Applicant before: Liu Baoquan |
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Effective date of registration: 20200312 Address after: 1500001 Room 101, building 10, no.1616, Chuangxin Road, Songbei District, Harbin City, Heilongjiang Province Applicant after: Harbin University of Technology International Stem Cell Engineering Research Institute Co.,Ltd. Address before: Room b1-5675, building 3, No.20 Yong'an Road, Shilong Economic Development Zone, Mentougou District, Beijing 102300 Applicant before: BEIJING HUANIU SHIJI BIOTECHNOLOGY RESEARCH INSTITUTE |
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