CN202482329U - NSCs (neural stem cell) continuous perfusion and cultivation system - Google Patents
NSCs (neural stem cell) continuous perfusion and cultivation system Download PDFInfo
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- CN202482329U CN202482329U CN 201220098403 CN201220098403U CN202482329U CN 202482329 U CN202482329 U CN 202482329U CN 201220098403 CN201220098403 CN 201220098403 CN 201220098403 U CN201220098403 U CN 201220098403U CN 202482329 U CN202482329 U CN 202482329U
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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
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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
Abstract
The utility model provides an NSCs (neural stem cell) continuous perfusion and cultivation system, which includes a perfusion and cultivation system and an intelligent monitoring system, wherein the perfusion and cultivation system includes a culture solution storer, a cell culture flask, a cell digestion flask, a waste liquid flask, a yield liquid flask, a speed-controlling fluid thruster, a speed controllable pipeline device, a fluid thruster and a filtering device, that are connected in the following ways: the culture solution storer is connected with the intelligent monitoring system through the speed controllable pipeline device via the speed-controlling fluid thruster, the intelligent monitoring system is connected with the cell culture flask through the speed controllable pipeline device via the speed-controlling fluid thruster, an outlet of the cell culture flask is connected with the filtering device through the speed controllable pipeline device, the fluid thruster is connected with the filtering device, and the intelligent monitoring system is connected with a bin inlet probe group and a bin outlet probe group at an inlet and the outlet of the cell culture flask respectively; all the devices are connected through the speed controllable pipeline system in a hermetic and sterile manner; the culture solution storer is used for storing sterile culture liquid for cell culture, and includes a shape-fixed sealed sterile tank system, a shape-fixed sealed sterile flask system or a shape-fixed sealed bag system; an outlet of the culture solution storer is connected with the speed controllable pipeline system in a hermetical and sterile manner; and the culture solution storer is provided with an air hole communicated with the atmosphere through a sterile filtration film filter.
Description
Technical field
The utility model relates to a kind of medical mechanism, particularly a kind of NSC continous pouring culture systems.
Background technology
Stem cell is meant the initiating cell with self and multidirectional differentiation potential, is the cells of origin of body, and can break up under given conditions becomes the cell that one or more constitute tissue or organ.
(Neural Stem Cell NSCs) is a kind of of stem cell to NSC, and NSC plays a significant role with repairing in the damaged nerve tissue at neurodevelopment.Neural stem cells transplantation is effective ways of repairing and replace impaired cerebral tissue, can rebuild part loop and function.NSC can be used as genophore in addition, is used for the gene therapy of intracranial tumors and other sacred disease, utilizes NSC as gene therapy vector, has remedied the some shortcomings of virus vector.In addition, NSC is observed the neural activity of some natural compounds and synthetic compound for judging that drug effect and drug toxicity etc. also have certain practical value as utilizing the Culture of neural stem cells technology, for development small molecules medicine is provided fundamental basis.
Along with the research to the NSC Application Areas is goed deep into, the culture of isolated of NSC also becomes one of focus of research instantly.At present the stripping technique of NSC has two kinds: 1. suspension culture partition method: the single cell suspension suspension culture in petridish after nervous tissue digestion is disperseed; Forming goes down to posterity behind the neural ball (about 200 microns of diameters) cultivates several times and separates NSC (the Experimental Neurology 156 that obtains; 71-83,1999).This method need rely on the neural ball of hand picking, very inconvenience in mass-producing is cultivated.2.FACS method: the specific antigens to NSC carries out fluorescent mark earlier, screens stem cell with precision instrument.Though this method can obtain the stem cell of higher degree; But stem cell there is certain infringement; And need expensive precision instrument (Proceeding of the National Academy of Sciences of the United States of America 97; 14720-14725,2000).
In sum, but study and develop high and stdn with low cost, the easy to operate control monitoring of a kind of separation purity, be convenient to the NSC continous pouring culture systems that robotization and mass-producing cultivate and become focus instantly.
The utility model content
The utility model discloses a kind of NSC continous pouring culture systems; To overcome the deficiency of existing artificial culture stem cell; Provide a kind of be used for the more stdn of laboratory or medical institutions, cell preparation applicable industry circle, rapidly and efficiently, NSC perfusion culture system that use range is wider, make that laboratory, hospital or factory can safe mass-producing ground high-level efficiency and prepare stem cell in high quality.
More specifically; The purpose of the utility model is, adopts the dynamic cellular culture technique, but a kind of device of continous pouring culture of neural stem cells neural is provided; While is connected with intelligence monitoring and controlling device, thereby constitutes the system of an intelligent control continous pouring culture of neural stem cells neural.
The system of a kind of continous pouring culture of neural stem cells neural that the utility model provides comprises perfusion culture system and intelligent monitor system (10), wherein:
Described perfused culture system comprises nutrient solution storer (1), Tissue Culture Flask (2), cell dissociation bottle (3), waste liquid bottle (4), results liquid bottle (5), controls fast liquid propeller (6), controlled fast plumbing installation (7), liquid propeller (8) and filtration unit (9); It is connected to: the nutrient solution storer is connected with intelligent monitor system through controlling fast liquid propeller through described controlled fast plumbing installation; Intelligent monitor system is connected with Tissue Culture Flask through controlling fast liquid propeller through controlled fast plumbing installation; The Tissue Culture Flask outlet connects filtration unit through controlled fast plumbing installation; Liquid propeller connects filtration unit, and intelligent monitor system links to each other with the probe group (14) that delivers from godown with the import of Tissue Culture Flask and the probe group (13) of putting in storage of outlet respectively; Connection between all devices connects and composes by controlled fast tubing system sealed, sterile; Described nutrient solution storer is in order to hold the aseptic culture fluid that supplies cell cultures; It comprises jar, bottle or the bag system of the sealed, sterile of solid shape, and its outlet is connected and is provided with through aseptic membrane filter with controlled fast tubing system sealed, sterile and is communicated with atmospheric pore;
Preferably, be provided with whisking appliance (11) in the described Tissue Culture Flask;
Preferably, being provided with an aperture in the described cell dissociation bottle is 6 microns filter membrane (12);
Preferably, described filtration unit is that an aperture is 200 microns a filtration unit, and liquid propeller can promote the neural ball of stem cell in the filtration unit to filter;
Preferably; Described intelligent monitor system is that the computer terminal of an automatic monitoring function and each sub-device that it is controlled comprise the system that its opening-closing valve constitutes; It can change different liquid storage interfaces as required under aseptic condition, so that different liqs flows into this perfused culture system; Its a plurality of indexs that can load said Tissue Culture Flask of insertion and the inner nutrient solution of said cell dissociation bottle are popped one's head in, and possess cultivation liquid level or nutrient solution weight in the monitoring reactor drum, detect cell concn, take a sample and detect, mix nutrient solution, control medium pH value, oxyty, temperature etc.;
Preferably, described Tissue Culture Flask is the sealed, sterile device, and upstream and downstream has access way respectively, and it has the structure of suitable NSC growth;
Preferably, said controlled fast tubing system is formed body by cell cultures silicone tube and fluid control speed thruster, and has the fast folder of the control of sliding between any two devices on the tubing system body or control fast valve, and the control speed of sliding is pressed from both sides the position or controlled fast valve and is positioned at the device exit;
Preferably, can the connect index probe of described intelligent monitor system of the controlled fast tubing system of the outlet of described connection culturing bottle and inlet flows into and flows out the index of the nutrient solution mixture of culturing bottle with monitoring;
Preferably, described fluid control speed thruster and controlled fast liquid propeller are conventional aseptic conveying liquid device.
The beneficial effect of the utility model is: after the utility model adopts aforesaid device to be combined into a system; Adopt the continuous perfused culture system of cell; Reactor drum and culturing bottle are combined, the mixing pre-treatment of acellular district substratum, the continuous perfusion of substratum mode of cellular regions are connected to a lasting flow process that unification is complete; Change liquid with the alternative manual work of the pipe switch mode of open and close different device, constantly import fresh reactant liquid, take away products of cellular metabolism; Is furnished with online Monitoring systems; Guarantee that each item index is stable, can greatly reduce material pollution, manpower transport, the reaction process stdn is controlled; And, in cell culture apparatus, set up the multilayer filter membrane to guarantee the normal effectively growth of NSC according to the suspension growth characteristic of NSC.The described device of the utility model is cultivated output height, easy to operate effective; Can greatly improve production efficiency, its preparation technology is simple, convenient quality control; Homogeneous, good NSC product be can efficiently obtain in a short time, scale operation and commercial applications are more suitable for; Use range is wide especially, therefore applies easily, can have a tremendous social and economic benefits in the short period of time.
Description of drawings
Fig. 1 is the synoptic diagram of system of the continous pouring culture of neural stem cells neural of the utility model.
Embodiment
The utility model has been studied existing medical mechanical skill, and a kind of system of continous pouring culture of neural stem cells neural is provided.
Extract the apparatus structure of stem cell for what more clearly describe the utility model and provided from placenta, below conjunction with figs. is specified.Accompanying drawing 1 is the synoptic diagram of system of the continous pouring culture of neural stem cells neural of the utility model.Provide shape and method of use and some experimental study contents of various culture vessels in following some experimental datas that provide and the following example by aforementioned utility model content; But should be appreciated that the utility model is not limited in listed research contents here; Should also be appreciated that term as used herein only is used to describe certain embodiments, and be not qualification the utility model.
Embodiment 1
The system of the continous pouring culture of neural stem cells neural that the utility model provides; Comprise perfusion culture system and intelligent monitor system (10); Wherein: described perfused culture system comprises nutrient solution storer (1), Tissue Culture Flask (2), cell dissociation bottle (3), waste liquid bottle (4), results liquid bottle (5), controls fast liquid propeller (6), controlled fast plumbing installation (7), liquid propeller (8) and filtration unit (9); It is connected to: the nutrient solution storer is connected with intelligent monitor system through controlling fast liquid propeller through described controlled fast plumbing installation; Intelligent monitor system is connected with Tissue Culture Flask through controlling fast liquid propeller through controlled fast plumbing installation; The Tissue Culture Flask outlet connects filtration unit through controlled fast plumbing installation; Liquid propeller connects filtration unit, and intelligent monitor system links to each other with the probe group (14) that delivers from godown with the import of Tissue Culture Flask and the probe group (13) of putting in storage of outlet respectively; Connection between all devices connects and composes by controlled fast tubing system sealed, sterile; Described nutrient solution storer is in order to hold the aseptic culture fluid that supplies cell cultures; It comprises jar, bottle or the bag system of the sealed, sterile of solid shape, and its outlet is connected and is provided with through aseptic membrane filter with controlled fast tubing system sealed, sterile and is communicated with atmospheric pore; Be provided with whisking appliance (11) in the described Tissue Culture Flask; Be provided with an aperture in the described cell dissociation bottle and be 6 microns filter membrane (12); Described filtration unit is that an aperture is 200 microns a filtration unit, and liquid propeller can promote the neural ball of stem cell in the filtration unit to filter; Described intelligent monitor system is that the computer terminal of an automatic monitoring function and each sub-device that it is controlled comprise the system that its opening-closing valve constitutes; It can change different liquid storage interfaces as required under aseptic condition, so that different liqs flows into this perfused culture system; Its a plurality of indexs that can load said Tissue Culture Flask of insertion and the inner nutrient solution of said cell dissociation bottle are popped one's head in, and possess monitoring reactor drum interior cultivation liquid level or nutrient solution weight size, detection cell concn, sampling detection, mix nutrient solution, control medium pH value, oxyty, temperature etc.; Described Tissue Culture Flask is the sealed, sterile device, and upstream and downstream has access way respectively, and it has the structure of suitable cell growth; Said controlled fast tubing system is formed body by cell cultures silicone tube and fluid control speed thruster, and has the fast folder of the control of sliding between any two devices on the tubing system body or control fast valve, and the control speed of sliding is pressed from both sides the position or controlled fast valve and is positioned at the device exit; Can the connect index probe of described intelligent monitor system of the controlled fast tubing system of the outlet of described connection culturing bottle and inlet flows into and flows out the index of the nutrient solution mixture of culturing bottle with monitoring; Described fluid control speed thruster and controlled fast liquid propeller are conventional aseptic conveying liquid device.
The system that the utility model provides in use; At first with the total system sterilization; The stem cell suspension that obtains is inoculated in the Tissue Culture Flask after the sterilization cultivates, during beginning perfusion culture liquid, open nutrient solution storer outlet conduit switch, intelligent monitor system pipe switch and open the fast liquid propeller of control; Nutrient solution in the nutrient solution storer is transported in the Tissue Culture Flask; Cell cultures situation in the Tissue Culture Flask will gather to intelligent monitor system simultaneously, and after the intelligent monitor system regulation and control, nutrient solution is in the condition that optimal cell is produced; Comprise the righttest oxyty, dissolve gas concentration lwevel, temperature, pH value etc., get into Tissue Culture Flask and carry out the continous pouring cell cultures; According to situation such as actual cell growths, the monitoring in real time in cell cultivation process of nutrient solution perfusion flow velocity is regulated with instant.Record cell growth each item index; When the stem cell nerve ball of some occurring; Open the Tissue Culture Flask gate out switch; Cell liquid is filtered by filter; Diameter moved into digest in the cell dissociation bottle greater than 200 microns neural ball break up, the cell dissociation bottle exports and is provided with an aperture and cell is discharged the entering waste liquid bottle less than 6 microns filter membrane in the lump when preventing to discharge digestive juice; Cell after breaing up reenters in the Tissue Culture Flask and cultivates, and opens the Tissue Culture Flask gate out switch when needing to digest the neural ball of breaing up once more until reappearing, and the stem cell suspension that obtains by Tissue Culture Flask is collected in the results liquid bottle.Through intelligent monitor system, can set switch and opening time length between each device of native system, make native system form an automatic successive culture of neural stem cells neural system, both can be, digest break up and cultivate simultaneously and carry out with filtration.In sum; The system design that the utility model provides a kind of NSC continous pouring to cultivate; Adopt the continuous perfused culture system of cell; Reactor drum and culturing bottle are combined, the mixing pre-treatment of acellular district substratum, the continuous perfusion of substratum mode of cellular regions are connected to a lasting flow process that unification is complete; Change liquid with the alternative manual work of the pipe switch mode of open and close different device, constantly import fresh reactant liquid, take away products of cellular metabolism; Is furnished with online Monitoring systems; Guarantee that each item index is stable, can greatly reduce material pollution, manpower transport, the reaction process stdn is controlled; And, in cell culture apparatus, set up the multilayer filter membrane to guarantee the normal effectively growth of NSC according to the suspension growth characteristic of NSC.The described device of the utility model is cultivated output height, easy to operate effective; Can greatly improve production efficiency, its preparation technology is simple, convenient quality control; Homogeneous, good NSC product be can efficiently obtain in a short time, scale operation and commercial applications are more suitable for; Use range is wide especially, therefore applies easily, can have a tremendous social and economic benefits in the short period of time.
Claims (9)
1. the system of a continous pouring culture of neural stem cells neural comprises perfusion culture system and intelligent monitor system (10), it is characterized in that:
Described perfused culture system comprises nutrient solution storer (1), Tissue Culture Flask (2), cell dissociation bottle (3), waste liquid bottle (4), results liquid bottle (5), controls fast liquid propeller (6), controlled fast plumbing installation (7), liquid propeller (8) and filtration unit (9); It is connected to: the nutrient solution storer is connected with intelligent monitor system through controlling fast liquid propeller through described controlled fast plumbing installation; Intelligent monitor system is connected with Tissue Culture Flask through controlling fast liquid propeller through controlled fast plumbing installation; The Tissue Culture Flask outlet connects filtration unit through controlled fast plumbing installation; Liquid propeller connects filtration unit, and intelligent monitor system links to each other with the probe group (14) that delivers from godown with the import of Tissue Culture Flask and the probe group (13) of putting in storage of outlet respectively; Connection between all devices connects and composes by controlled fast tubing system sealed, sterile; Described nutrient solution storer is in order to hold the aseptic culture fluid that supplies cell cultures; It comprises jar, bottle or the bag system of the sealed, sterile of solid shape, and its outlet is connected and is provided with through aseptic membrane filter with controlled fast tubing system sealed, sterile and is communicated with atmospheric pore.
2. the system of continous pouring culture of neural stem cells neural as claimed in claim 1 is characterized in that: be provided with whisking appliance (11) in the described Tissue Culture Flask.
3. the system of continous pouring culture of neural stem cells neural as claimed in claim 1 is characterized in that: be provided with an aperture in the described cell dissociation bottle and be 6 microns filter membrane (12).
4. the system of continous pouring culture of neural stem cells neural as claimed in claim 1 is characterized in that: described filtration unit is that an aperture is 200 microns a filtration unit.
5. the system of continous pouring culture of neural stem cells neural as claimed in claim 1; It is characterized in that: described intelligent monitor system is that the computer terminal of an automatic monitoring function and each sub-device that it is controlled comprise the system that its opening-closing valve constitutes; It can change different liquid storage interfaces as required under aseptic condition, so that different liqs flows into this perfused culture system; It can load a plurality of index probes that insert said Tissue Culture Flask and the inner nutrient solution of said cell dissociation bottle.
6. the system of continous pouring culture of neural stem cells neural as claimed in claim 1 is characterized in that: described Tissue Culture Flask is the sealed, sterile device, and upstream and downstream has access way respectively, and it has the structure of suitable NSC growth.
7. the system of continous pouring culture of neural stem cells neural as claimed in claim 1; It is characterized in that: said controlled fast tubing system is formed body by cell cultures silicone tube and fluid control speed thruster; And have the fast folder of the control of sliding between any two devices on the tubing system body or control fast valve, the control speed of sliding is pressed from both sides the position or is controlled fast valve and is positioned at the device exit.
8. the system of continous pouring culture of neural stem cells neural as claimed in claim 1; It is characterized in that: can the connect index probe of described intelligent monitor system of the controlled fast tubing system of the outlet of described connection culturing bottle and inlet flows into and flows out the index of the nutrient solution mixture of culturing bottle with monitoring.
9. the system of continous pouring culture of neural stem cells neural as claimed in claim 1 is characterized in that: described fluid control speed thruster and controlled fast liquid propeller are conventional aseptic conveying liquid device.
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| CN 201220098403 CN202482329U (en) | 2012-03-15 | 2012-03-15 | NSCs (neural stem cell) continuous perfusion and cultivation system |
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| CN 201220098403 CN202482329U (en) | 2012-03-15 | 2012-03-15 | NSCs (neural stem cell) continuous perfusion and cultivation system |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103484368A (en) * | 2013-09-06 | 2014-01-01 | 北京科兴生物制品有限公司 | Operation pipe system for cell factory and application thereof |
| CN103480276A (en) * | 2013-09-06 | 2014-01-01 | 北京科兴生物制品有限公司 | Closed ultrafiltration pipeline system and application thereof |
| CN103484365A (en) * | 2013-08-30 | 2014-01-01 | 中国人民解放军南京军区福州总医院 | Device and method for obtaining stem cells from tissues |
-
2012
- 2012-03-15 CN CN 201220098403 patent/CN202482329U/en not_active Expired - Lifetime
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103484365A (en) * | 2013-08-30 | 2014-01-01 | 中国人民解放军南京军区福州总医院 | Device and method for obtaining stem cells from tissues |
| CN103484365B (en) * | 2013-08-30 | 2015-09-16 | 中国人民解放军南京军区福州总医院 | A kind of devices and methods therefor obtaining stem cell from tissue |
| CN103484368A (en) * | 2013-09-06 | 2014-01-01 | 北京科兴生物制品有限公司 | Operation pipe system for cell factory and application thereof |
| CN103480276A (en) * | 2013-09-06 | 2014-01-01 | 北京科兴生物制品有限公司 | Closed ultrafiltration pipeline system and application thereof |
| CN103480276B (en) * | 2013-09-06 | 2015-09-02 | 北京科兴生物制品有限公司 | A kind of closed ultrafiltration pipe-line system and application thereof |
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| C56 | Change in the name or address of the patentee |
Owner name: SHANGHAI ANGECON BIO-TECHNOLOGY CO., LTD. Free format text: FORMER NAME: SHANGHAI ANGECON BIOTECHNOLOGY CO., LTD. |
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| CP03 | Change of name, title or address |
Address after: 201318 Shanghai City, Pudong New Area Road No. 588 building 3 indigo Patentee after: SHANGHAI ANGECON BIOTECHNOLOGY CO., LTD. Address before: 201203 Shanghai city Pudong New Area Zhangjiang High Tech Park of Shanghai Cailun Road Lane 720 Building No. 2 room 504 Patentee before: Shanghai Angecon Biotechnology Co., Ltd. |
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Granted publication date: 20121010 |