CN2923715Y - Automatic candidate steam cell culturing system - Google Patents
Automatic candidate steam cell culturing system Download PDFInfo
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- CN2923715Y CN2923715Y CN 200520043614 CN200520043614U CN2923715Y CN 2923715 Y CN2923715 Y CN 2923715Y CN 200520043614 CN200520043614 CN 200520043614 CN 200520043614 U CN200520043614 U CN 200520043614U CN 2923715 Y CN2923715 Y CN 2923715Y
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- chamber
- stem cell
- gas
- culture
- stem cells
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Abstract
The utility model relates to a biological cell culture system, especially relates to a vitro amplification culture device for human stem cells or/and human hematopoiesis stem cells, which is a device of auto-culture and amplification of stem cells and hematopoiesis stem cells concretely. In the device, each kind of stem cells and hematopoiesis stem cells of various differentiations are cultured synchronously, the nutrient composition of the automatic adjustment and control culture media keeps in the suitable level with growth factor, and the product is metabolized in the auto-excretion culture. The device and means of vitro synchronization culture of a plurality of stem cells and hematopoiesis stem cells can be used in clinical biological cells treatment and hematopoiesis stem cells transplantation.
Description
Affiliated technical field
The utility model relates to a kind of biomass cells culture systems, relate in particular to a kind of human stem cell or/and human hematopoietic stem amplification in vitro culture apparatus, more particularly, this is the device that a kind of stem cell and hemopoietic stem cell are cultivated and increased automatically, in this device, the stem cell of various different differentiation degrees and hemopoietic stem cell are by synchronized culture, automatically regulate and control the nutrition and the somatomedin of cultivating in the media and remain on proper level, automatically drain meta-bolites in the cultivation, this apparatus and method at external synchronized culture broad variety stem cell and hemopoietic stem cell can be applied to clinical biological cell therapeutical and transplantation of hematopoietic stem cell.
Background technology
Research and development are just thirsted for by bio-science man for a long time in the system of external mass production human bone marrow and other hemopoietic stem cells, because such system can not only satisfy extensive and diversified clinical needs, simultaneously also can provide great help for scientific research and applied research, its application is as follows:
The elementary dynamics research of 1 hemopoietic stem cell differentiation
2 improve from body or simplified marrow transplanting
3 emptyings or remove any cell of occurring of not wishing in bone marrow transplantation are as T cell or other fatal cells
The gene therapy of 4 blood cell systems
5 improve miscellaneous tumour cell in human body autologous peripheral blood stem cell transplantation, the purification stem cell
6 improve navel blood stem cell transplanting, amplification navel blood stem cell quantity, to satisfy adult's oncotherapy
7 are used to induce the differentiation dendritic cell, carry out immunotherapy of tumors and the tumor vaccine treatment of dendritic cell for the basis
8 a large amount of human hematopoietic stems that produce are as CD34
+Cell uses for clinical or research
All kinds of mature blood cells of 9 a large amount of generations are as red corpuscle, thrombocyte.
Although long-term hemopoietic stem cell is cultivated (" Long-term Hematopoietic culture " " LTHBMC ") and is suggested in the later stage eighties, recent development has significantly improved original method, and has produced revolutionary change.
Yet these improvement remain the subclinical quantity medullary cell implemented or the method for other hemopoietic stem cells under the tissue culturing equipment of standard laboratory scale.
Verified these the recent improvement of a series of research are successful, the function of supported matrix cellular layer is shown, cultivate the filling rate of media or the exchange sequence of cultivation media and be subjected to specific effect, metabolic function, grow up, the more important thing is that perhaps the secretion of growth factor all has been proved to be the cultivation media exchange rate influence that is subjected to normal human's marrow fibroblast.
In the vitro culture environment, stroma cell is supported the ability of human hematopoietic stem to be proved to be can be cultivated exchange rate by media apace to improve, in exchange of fast culture media and highdensity cell, LTHBMC can support a large amount of stable immature cells that produce in the hemopoietic stem cell to reach for 20 weeks, these evidences that formed prove, under these culture environment, can stablize a large amount of generation hemopoietic stem cells.
Correctly adopt and further to improve these cultured method by the dissolved cell growth factor.
Noticeable and eloquent demand is present among the method for these propositions, promptly has only to develop the composing method that can implement clinical quantity and device could be realized above-mentioned required application.
In the bio-reactor design in the past, although being used as important aspect, the cell harvesting program considers, but the method that proposes remains the structure of wide open type, when harvested cell, must stop culturing process, and must expose the cultivation chamber in atmosphere, this structure is easy to constitute the pollution of cultivating chamber.
Further, bio-reactor need keep a kind of equilibrium state of inside, especially in the initiating cell culturing process of complexity, human stem cell or mankind hemopoietic stem cell are to their power very responsive (being gas filling rate/nutritive medium filling rate/somatomedin filling rate), also very responsive to the supply and the removal of above-mentioned substance simultaneously, chemical environment to them is responsive equally, in the present bio-reactor, can satisfy above-mentioned requirements without any design, thereby keep cultivating homeostasis and stable, especially in the initiating cell culturing process.
Also fail to propose a kind of method harvested cell under the state that does not interrupt cultivating in the present design, equally also do not have a kind of design can keep cultivating interior environment and keep balance.Stable gas is provided, the liquid filling rate, the method that possesses above-mentioned two kinds of advantages is not simultaneously more found.
Summary of the invention
The purpose of this utility model provides a kind of totally enclosed, HT rate, high media cultivation exchange rate, the hematopoietic stem cell culturing apparatus of harvested cell under the state that does not interrupt cultivating, for environment in cultivating keeps balance, provide stable gas and liquid filling rate.
In order to realize the purpose of this utility model, the utility model proposes a kind of novel culture apparatus.Its system constitutes: by cultivating chamber, upper and lower two-layer gas chamber and all around four abandon the sap cavity chamber, monitoring and Controlling System automatically, gangway and manually-operated gate thereof, magnetic valve, peristaltic pump, gas supply system etc. are partly formed.
● cultivate chamber
■ is cell growth place, cultivate the upper and lower two-layer gas semi-permeable membranes that is of chamber, the semi-permeable membranes surface is handled (corona treatment) thereby stronger cell adhesion characteristic is arranged through electrical spark, ventilative semi-permeable membranes leans against sieve tray (big metering-orifice is arranged on the wooden partition) can play the support semi-permeable membranes, thereby the function of enhancing pressure resistivity, but again because its sieve aperture structure, can extensively carry out gaseous interchange again, isolate with the good silicone rubber gasket of nontoxic element, biocompatibility between sieve tray and the gas semi-permeable membranes and fixing.
■ cultivates chamber four sidewalls and is supermicro filtration membrane (NWCO is 50,000), can overflow products of cellular metabolism and part moisture content, but can keep the most nutrition of all cells, and product in the cell, when being used for continuing perfusion, carrying out nutrition and exchanges.
■ cultivates the chamber right side wall and leaves import and export, and for the input cell nutrient solution, the sampling monitoring continues purposes such as perfusion and harvested cell.
■ cultivates chamber size and can design different size, from 50ml～250ml according to clinical application.
It is flats that ■ cultivates chamber, and width and length are tens of times of height, helps gas and fills the air and save nutritive medium, nutrient solution (highly being no more than 4mm).
The ■ culture chamber is indoor PH, glucose (dissolved oxygen) transmitter such as (DO), monitoring in time, control culture condition and culture environment.
■ cultivates the chamber below the electronic weighing scale that weighs, and measures weight (weight of culture vessel is constant number, can judge indirectly culture chamber indoor cultivation liquid how much) at any time.
It is totally-enclosed system that ■ cultivates chamber, only can see through semi-permeable membranes and carry out gaseous interchange, and carry out the nutrition exchange by supermicro filtration membrane, and each quasi-microorganism can't penetrate.
● gas chamber
■ is cultivating chamber upper and lower two-layer (top layer and bottom), be gas chamber, the bottom of gas chamber (upper strata) or top (lower floor), be sieve tray, gas chamber is integrated, its sieve tray is connected with semi-permeable membranes and sidewall (culture chamber sidewall) (being connected by long screw) by silicon rubber (upper and lower two-layer).
There are gas inlet and outlet in ■ gas chamber, and blended oxygen, carbonic acid gas is by inlet inflow gas chamber, then wherein partly by the disperse of gas semi-permeable membranes to culture chamber, all the other partly flow out by outlet.
In the ■ gas chamber O is arranged
2And CO
2Transmitter continues to monitor the O of gas chamber
2And CO
2Content, and regulate O by varitrol
2And CO
2Content.
● abandon sap cavity chamber (nutrition exchange sap cavity chamber)
■ is positioned at and cultivates four sidewalls of chamber, the sidewall sieve tray is close to supermicro filtration membrane, and (supermicro filtration membrane tackiness agent, inorganic nontoxic binding material stick on supermicro filtration membrane on the sidewall sieve tray, the also screw that can make with non-toxic plastic, with nontoxic silicone rubber gasket fixedly supermicro filtration membrane on sieve tray).
■ abandons the sap cavity chamber has sidewall (equidirectional with the culture chamber chamber inlet) that an outlet (being positioned at the side plate bottom) is arranged, and nutrition exchange liquid can flow out from outlet.
The bottom plate had certain slope (obliquity) around ■ abandoned the sap cavity chamber, tilted to Way out, so that nutrition exchange liquid flows out.
● cultivate the chamber gangway
■ is positioned at and cultivates chamber right side wall, central part.
■ is used to cultivate the perfusion and the extraction of chamber contents.
The manually-operated gate of one two direction of ■ gangway place's device, all sealings, system closing (the cap sealing is arranged).
Current downwards: (the cap sealing is arranged, and cap is disposable use)
Be used for: import compositions such as micro-cell growth factor with syringe, in cultivation, extract partly nutrient solution observation of cell growing state, harvested cell (being connected) with TFF automated cell harvesting system.
Upwards current: (there is electromagnetic valve control the pipeline top)
Be used for: perfusion inoculation culture cell, continue the perfusion nutritive medium in the cultivation, the perfusion damping fluid is used for cell harvesting.
● abandon the liquid chamber outlet
■ is positioned at and abandons the liquid chamber right side wall near bottom position, (equidirectional with the culture chamber chamber inlet).
■ is used to dump nutrition exchange liquid.
Its outlet of ■ is abandoned bottom, sap cavity chamber and is tilted to Way out for abandoning sap cavity chamber extreme lower position, and nutrition exchange liquid flows out outlet by action of gravity.
Be connected on its outlet conduit of ■ and abandon the liquid bag, outlet conduit has magnetic valve or manually-operated gate, can seal.
The ■ outlet conduit links to each other with peristaltic pump, can aspirate in case of necessity.
● the gas chamber gangway
■ gas chamber inlet is positioned at gas chamber's right side wall, and gas chamber's outlet is positioned at gas chamber's left side wall.
All there is electromagnetic valve control at ■ gas chamber gangway pipeline place, can control its open and close and flow thereof and flow velocity.
The gas-filtering device of Pall company, filtering microorganism all are equipped with in the gangway in ■ gas chamber.
● multi-parameter monitoring and warning system
In culturing process, monitor multiple parameters at any time.
■ O
2And CO
2Flow velocity, pressure and flow.At O
2And CO
2In four input channels rate of flow meter is housed, digital monitoring gas flow, flow velocity are set warning device.
■ O
2And CO
2Content.O is housed in gas chamber
2And CO
2Transmitter can be monitored O in the gas chamber at any time
2And CO
2Content, and warning and adjusting automatically are set.
■ PH monitoring.The PH electrode is contained in the culturing room, monitors the pH value of nutritive medium at any time.
The ■ glucose monitoring.It is indoor that employing One Touch glucose monitoring transmitter is contained in culture chamber.
The monitoring of ■ cell.Adopt (BDS) automatic bacteria proofing unit regularly to detect bacterium in the nutrient solution (drawing liquid microscopy simultaneously).
The groundwater increment monitoring of ■ nutritive medium.Below culture apparatus, install a digitizing weight meter, measuring apparatus weight at any time, indirect calculation nutritive medium groundwater increment.
■ automatic filling flow monitoring.Under meter is housed on inlet duct can monitors irrigation flow, the speed digitizing of peristaltic pump simultaneously shows its rotating speed.
The above parameters monitoring of ■ is digital monitoring.In time write down all parameters with computer.
■ can preset various indexs according to cultivating requirement, in case touch these indexs, can report to the police sound and light alarm automatically.
■ links to each other with walky-talky, if any any alarm, and the automatic dialing of meeting, and on assigned telephone number, leave a message.(or staying information on Palm type Cell phone device) can in time notify laboratory worker outside in time to go back to the laboratory.
● automatic control system
The ■ automatic temperature control system.Automatic constant-temperature is controlled at the culture temperature of setting, as the warning automatically that changes.
■ O
2And CO
2Gas is controlled automatically.According to O
2And CO
2The transmitter monitoring result is regulated automatically.
The ■ nutritive medium continues automatic filling control.Digitizing shows and the peristaltic pump of control is controlled automatically, regulates control according to the groundwater increment feedback of automatic monitoring.
The automatic control of ■ can be adopted micro computer process chip (DSP) or adopt desktop computer (touch screen computer) control.(automatic control system can be purchased finished product (bioprocess controller), homemade or import).
■ shows.Can show and the touch screen monitor demonstration with large-screen lc.
● peristaltic pump.Adopt the Digital Control peristaltic pump.Digitizing demonstration and Digital Control, governing speed automatically.
● magnetic valve.Adopt the electromagnetism squeeze valve, by automatic controlling system.
● the automated cell harvesting system.Adopt TFF automated cell harvesting system to come harvested cell.
● according to the material difference that is adopted, can be divided into:
The disposable using appts of ■.Material therefor is a non-toxic duroplasts, mould compacting, disposable use.Sterilization (throwing aside after the use) is packaged in standby in the sterile packed (disinfection with high pressure steam) after the sterilization in advance, and the gas semi-permeable membranes is connected employing alite paste bonding method with supermicro filtration membrane with the plastics sieve tray.
■ reuses device.Material therefor is metallic substance (as stainless steel etc.).The gas semi-permeable membranes adopts silicon rubber pad+screw to be connected with supermicro filtration membrane.Need device gas semi-permeable membranes and supermicro filtration membrane before using, and need disinfection with high pressure steam before use.
● be applicable to the various kinds of cell cultivation, especially clinical biological cell therapeutical is used.
■ hemopoietic stem cell vitro culture, amplification (marrow, peripheral blood, bleeding of the umbilicus)
■ dendritic cell amplification in vitro is induced (marrow, peripheral blood, bleeding of the umbilicus, CD34
+Source or monocyte source)
■ autologous tumor vaccine production.
■ filters immunocyte treatment (cell therapies such as LAK, TIL, CIK, TCK) from body or allosome
The ■ Monoclonal Antibody; Thrombocyte, red corpuscle amplification in vitro.
● built-up type multi-unit cells culture systems.
A plurality of unit cell culture system can combine built-up type multi-unit cells culture systems, can be four unit-combination type cell culture systems, eight unit-combination type cell culture systems, 16 unit-combination type cell culture systems, enjoy a cover automatic control system jointly, one group of O
2, CO
2Transmitter and Controlling System.One group of automatic continuous filling type system.The unit cost of culture apparatus significantly reduces, and efficient improves, and help extensive regional laboratory and use, or large hospital uses.
The beneficial effects of the utility model provide a kind of hemopoietic stem cell culture systems of simple in structure, practical and convenient, automatic control, greatly simplify hematopoietic stem cell population process in the clinical application, reduce the pollution rate of sample, improve transplanting succeed rate.
Description of drawings
The utility model is described in further detail below in conjunction with accompanying drawing
Fig. 1 is the utility model device sectional view
Fig. 2 is gas chamber's synoptic diagram
Fig. 3 is apparatus of the present invention vertical views
Fig. 4 is the side plate synoptic diagram
Fig. 5 is apparatus of the present invention connection diagrams.
Fig. 6 is a built-up type multi-unit cells culture systems synoptic diagram.
Embodiment
101-damping fluid among Fig. 1,102-nutritive medium (continuing perfusion), the 103-inoculating cell, 104-peristaltic pump (band cap, manual bidirectional regulated valve), 105-injects results/micro-injection/liquid pumping hole, 106-O
2CO
2Inlet, 107-O
2CO
2Transmitter, 108-gas chamber, 109-O
2CO
2Outlet, 110-abandons sap cavity, the 111-sieve tray, the 112-supermicro filtration membrane, 113-cultivates chamber, 114-PH transmitter, 115-glucose sensor, 116-gas chamber, 117-sieve tray, 118-gas semi-permeable membranes.
201-gas chamber among Fig. 2,202-screw (band inside screw), 203-sieve plate hole.
304-liquid inlet among Fig. 3,305-O
2CO
2Inlet, 306-C
2CO
2Transmitter, 307-gas semi-permeable membranes, 308-gas chamber, the 309-O2CO2 outlet, 310-abandons liquid chamber, the 311-sieve plate, 312-supermicro filtration membrane, 313-PH transmitter, 314-glucose sensor, 316-cultivates chamber, 317-gas chamber, and 318-abandons liquid outlet, 319-sieve plate.
401-screw among Fig. 4 (band internal thread), 402-sidewall sieve tray, 403-supermicro filtration membrane.
501-gas chamber among Fig. 5,502-air chamber wall, 503-silicone rubber gasket, 504-gas semi-permeable membranes, 505-silicone rubber gasket, the 506-sidewall, 507-sidewall sieve tray, 508-sieve tray, 509-gas chamber, 510-air chamber wall, 511-gas semi-permeable membranes, 512-packing ring, 513-sidewall.
601-abandons liquid among Fig. 6, the 602-nutritive medium, and 603-gas inlet pipeline, 604-gas mixes and gas control equipment, 605-O
2And CO
2Gas cylinder, 606-gas sensor, 607-air filter, 608-pneumatic outlet pipeline.
Claims (5)
1. automatic hemopoietic stem cell culture systems, it is characterized in that: the hemopoietic stem cell culture systems is by cultivating chamber automatically, upper and lower two-layer gas chamber and all around four abandon the sap cavity chamber, automatically monitor and Controlling System, gangway and manually-operated gate thereof, magnetic valve, peristaltic pump, gas supply system is formed.
2. according to the described automatic hemopoietic stem cell culture systems of claim 1, it is characterized in that: cultivate the upper and lower two-layer gas semi-permeable membranes that is of chamber, handle through electrical spark on the semi-permeable membranes surface, ventilative semi-permeable membranes leans against sieve tray, isolates with the good silicone rubber gasket of biocompatibility between sieve tray and the gas semi-permeable membranes and fixing.
3. according to the described automatic hemopoietic stem cell culture systems of claim 1, it is characterized in that: abandon the sap cavity chamber and be positioned at four sidewalls of cultivation chamber, the sidewall sieve tray is close to supermicro filtration membrane, abandon the sap cavity chamber sidewall is arranged, being positioned at side plate bottom has an outlet, and nutrition exchange liquid flows out from outlet, abandon the sap cavity chamber around the bottom plate certain slope is arranged, tilt to Way out, so that nutrition exchange liquid flows out.
4. according to the described automatic hemopoietic stem cell culture systems of claim 1, it is characterized in that: the multi-parameter monitoring of setting and alarm unit can be monitored multiple parameters at any time in culturing process, at O
2And CO
2In four input channels rate of flow meter is housed, O is housed in gas chamber
2And CO
2Transmitter, dress PH electrode at the indoor dress glucose monitoring of culture chamber transmitter, below culture apparatus, installs a digitizing weight meter in culturing room, and under meter is housed on inlet duct.
5. according to the described automatic hemopoietic stem cell culture systems of claim 1, it is characterized in that: control automatically and adopt the micro computer process chip or adopt desktop computer control, show and the touch screen monitor demonstration with large-screen lc.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200520043614 CN2923715Y (en) | 2005-07-21 | 2005-07-21 | Automatic candidate steam cell culturing system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200520043614 CN2923715Y (en) | 2005-07-21 | 2005-07-21 | Automatic candidate steam cell culturing system |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN2923715Y true CN2923715Y (en) | 2007-07-18 |
Family
ID=38260144
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200520043614 Expired - Fee Related CN2923715Y (en) | 2005-07-21 | 2005-07-21 | Automatic candidate steam cell culturing system |
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| Country | Link |
|---|---|
| CN (1) | CN2923715Y (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102174395A (en) * | 2011-01-30 | 2011-09-07 | 中国科学院广州生物医药与健康研究院 | Automation augmentation and culture system of induced pluripotent stem cells |
| CN102199535A (en) * | 2010-03-23 | 2011-09-28 | 上海坤巨科技发展有限公司 | Device and method for noninvasive continuous monitoring of quantity or concentration of dynamic cells |
| CN102199536A (en) * | 2010-03-23 | 2011-09-28 | 上海坤巨科技发展有限公司 | Apparatus and method for monitoring cell quantity or cell proportion during dynamic stem cell differentiation |
| CN105316227A (en) * | 2014-07-28 | 2016-02-10 | 温州医科大学附属第二医院 | Multifunctional cell simulation cabin |
| CN113423809A (en) * | 2018-11-08 | 2021-09-21 | 阿格拉瑞斯有限公司 | Cell culture system and method |
-
2005
- 2005-07-21 CN CN 200520043614 patent/CN2923715Y/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102199535A (en) * | 2010-03-23 | 2011-09-28 | 上海坤巨科技发展有限公司 | Device and method for noninvasive continuous monitoring of quantity or concentration of dynamic cells |
| CN102199536A (en) * | 2010-03-23 | 2011-09-28 | 上海坤巨科技发展有限公司 | Apparatus and method for monitoring cell quantity or cell proportion during dynamic stem cell differentiation |
| CN102174395A (en) * | 2011-01-30 | 2011-09-07 | 中国科学院广州生物医药与健康研究院 | Automation augmentation and culture system of induced pluripotent stem cells |
| CN102174395B (en) * | 2011-01-30 | 2013-08-21 | 中国科学院广州生物医药与健康研究院 | Automation augmentation and culture system of induced pluripotent stem cells |
| CN105316227A (en) * | 2014-07-28 | 2016-02-10 | 温州医科大学附属第二医院 | Multifunctional cell simulation cabin |
| CN113423809A (en) * | 2018-11-08 | 2021-09-21 | 阿格拉瑞斯有限公司 | Cell culture system and method |
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|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C19 | Lapse of patent right due to non-payment of the annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |