CN104403946A - Stem cell culture perfusion device - Google Patents
Stem cell culture perfusion device Download PDFInfo
- Publication number
- CN104403946A CN104403946A CN201410812200.7A CN201410812200A CN104403946A CN 104403946 A CN104403946 A CN 104403946A CN 201410812200 A CN201410812200 A CN 201410812200A CN 104403946 A CN104403946 A CN 104403946A
- Authority
- CN
- China
- Prior art keywords
- cell
- perfusion
- cell culture
- peristaltic pump
- stem cell
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- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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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
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
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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
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
- C12M29/04—Filters; Permeable or porous membranes or plates, e.g. dialysis
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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
- C12M41/26—Means for regulation, monitoring, measurement or control, e.g. flow regulation of pH
Abstract
The invention provides a stem cell culture perfusion device which comprises a perfusion bottle, a feeder layer cell culture unit, a cell perfusion tank and a waste liquid bottle, wherein the opening of the to-be-cultured cell perfusion bottle is provided with a first filter head and an input tube; the input tube is provided with a first controller capable of displaying pH value and controlling temperature; the tail end of the input tube is immersed in the feeder layer cell culture unit; a first connecting tube, which is connected through the feeder layer cell culture unit of which the substrate is provided with a semi-permeable membrane, is provided with a second controller and a first peristaltic pump; both ends of the cell perfusion tank are respectively connected with a second connecting tube and a third connecting tube; the other end of the second connecting tube is connected with the first peristaltic pump; the other end of the third connecting tube is connected with the a second peristaltic pump; the opening of the waste liquid bottle is provided with an output tube and a second filter head; and the other end of the output tube is connected with the second peristaltic pump. Under the actions of the first controller and second controller, the temperature and pH value variations can be observed, the rate of the peristaltic pumps can be controlled, and error values caused by manual observation and recording can be avoided; and the equipment output of the cell perfusion tank can enhance the stem cell culture rate.
Description
Technical field
The invention belongs to a kind of perfusion device, particularly relate to a kind of for stem cell culture perfusing device.
Background technology
In embryonic stem cell vitro culture, except normal substratum, need first to cultivate the special cell of one deck (as mouse embryo fibroblast-mouse embryonic fibroblast MEF cell etc.), and then inoculate the stem cell that will test thereon.These special cells are exactly feeder layer cells, normally become fiber-like cell, these cells can secrete the necessary somatomedin of some the unknowns as leukaemia inhibitory factor (leukemia inhibitoryfactor, LIF), STEM CELL FACTOR (stem cell factor, and Prostatropin (basic Fibroblast growthfactor SCF), bFGF) etc., provide embryonic stem cell, induced multi-potent stem cells better nutrition.The survival of these factor pair stem cells and propagation are required, and its quantity not only can be made to increase, and relevant with stem cell differentiation in vivo and in vitro.Experiment proves, the effect of feeder layer cells is not only create some cytokines, and in most cells growth and atomization, the adhesion of cell has all played considerable effect.ES cell can not be survived as completely independently cell, is always associated with the cell of surrounding, and the adhesion receptors in intracellular signaling regulates the motion of cell, propagation and differentiation.Usually under embryonic stem cell/induced multi-potent stem cells culture system, feeder layer cells and stem cell are in Dual culture state, and both cannot be separated thoroughly.If adopt the mode of feeder layer and stem cell difference static cultivation, cannot in real time the factor that feeder layer cells is secreted be sent in stem cell culture system, and operating process is many, is scarcely used.Tradition Dual culture mode affects stem cell purity, and in stem cell application etc., serve very large restriction, the speed of reaction improving culture apparatus is undoubtedly the common objective of those skilled in the art.
Summary of the invention
The problem to be solved in the present invention is to provide stem cell culture perfusing device, the inconvenient Real Time Observation of the conventional perfusion device used, and cultivates speed simultaneously and has much room for improvement.
For solving the problems of the technologies described above, the technical solution used in the present invention is:
Stem cell culture perfusing device, comprise perfusion bottle, feeder layer cells incubator, vacuum culture device and waste liquid bottle, described perfusion bottle bottle mouth position arranges the first filter and input tube, described first filter can make the pressure in described perfusion bottle identical with the external world, and the bacterial contamination that can completely cut off in outside air, described input tube is arranged the first controller of a control temperature and pH value, described input tube tail end immerses feeder layer cells incubator, the first pipe connecting that the feeder layer cells incubator arranging semi-permeable membranes by substrate connects arranges a second controller and the first peristaltic pump, guarantee that the cell in feeder layer cells incubator can carry out cell cultures at the temperature adapting to cell proliferation and flow velocity, bottom described cell perfusion groove inner chamber body, the sterile-processed absorbent cotton of a regular shape is set, absorbent cotton fits in bottom cell perfusion groove inner chamber body, described absorbent cotton top arranges a reticulated structure, reticulated structure top arranges the perfusion groove support portion of placing cell, by absorbent cotton and reticulated structure acting in conjunction, the response location of perfusion groove support portion can be raised, make the fluidizing air of the nutritious material of the abundant contact zones of cell, absorbent cotton and reticulated structure all directly can not contact with cell culture fluid, with reticulated structure, absorbent cotton can also guarantee that perfusion groove support portion does not directly contact with bottom cell perfusion groove inner chamber body the effect playing insulation simultaneously, described cell perfusion groove bottom arranges a bracing frame, guarantee that cell perfusion groove obtains firm support in certain position aloft, described cell perfusion groove two ends connect the second pipe connecting and the 3rd pipe connecting respectively, the described second pipe connecting the other end is connected with described first peristaltic pump, the described 3rd pipe connecting the other end is connected with described second peristaltic pump, described waste liquid bottle bottleneck arranges output tube and the second filter, described second filter can make the pressure in described waste liquid bottle identical with the external world, and the bacterial contamination that can completely cut off in outside air, the described output tube the other end connects described second peristaltic pump.
Further, bottom described feeder layer cells incubator, a Tissue Culture Plate is set, and described semi-permeable membranes is positioned on described Tissue Culture Plate.
Further, described cell perfusion groove two ends arrange the semi-permeable membranes preventing culturing cell from passing, but semi-permeable membranes can make liquid freely pass through.
Further, in the middle part of described cell perfusion groove, an annular transparent hole is set, facilitates observation of cell perfusion groove inner cell to breed situation.
The advantage that the present invention has and positively effect are: a kind of stem cell culture perfusing device of the present invention, this device passes through the effect of the first controller and second controller, under observable differing temps, pH value is changed and is judged the flow velocity of peristaltic pump by different pH value, can avoid again producing error value because of manual measurement simultaneously, the effect of the semi-permeable membranes at two peristaltic pumps, absorbent cotton and cancellated uses and cell perfusion groove two ends, improve the cultivation speed of stem cell, cell perfusion groove can also be made to play heat insulation effect.
Accompanying drawing explanation
Fig. 1 is schematic diagram of the present invention
In figure:
1-perfusion bottle 2-first filter 3-input tube 4-first controller
5-feeder layer cells incubator 6-first pipe connecting 7-second controller
8-first peristaltic pump 9-second pipe connecting 10-cell perfusion groove
11-the 3rd pipe connecting 12-second peristaltic pump 13-output tube
14-second filter 15-waste liquid bottle 16-bracing frame
Embodiment
Below in conjunction with accompanying drawing, specific embodiments of the invention are elaborated.
As shown in Figure 1, stem cell culture perfusing device, comprise perfusion bottle 1, feeder layer cells incubator 5, cell perfusion groove 10 and waste liquid bottle 15, perfusion bottle 1 bottle mouth position arranges the first filter 2 and input tube 3, first filter 2 can make the pressure in perfusion bottle 1 identical with the external world, and the bacterial contamination that can completely cut off in outside air, input tube 3 is arranged the first controller 4 of a control temperature and pH value, input tube 3 tail end immerses feeder layer cells incubator 5, the first pipe connecting 6 that the feeder layer cells incubator 5 arranging semi-permeable membranes by substrate connects arranges second controller 7 and the first peristaltic pump 8, guarantee that the cell in feeder layer cells incubator 5 can carry out cell cultures at the temperature adapting to cell proliferation and flow velocity, bottom cell perfusion groove 10 inner chamber body, the sterile-processed absorbent cotton of a regular shape is set, absorbent cotton fits in bottom cell perfusion groove 10 inner chamber body, absorbent cotton top arranges a reticulated structure, reticulated structure top arranges the perfusion groove support portion of placing cell, by absorbent cotton and reticulated structure acting in conjunction, the response location of perfusion groove support portion can be raised, make the fluidizing air of the nutritious material of the abundant contact zones of cell, improve the cultivation speed of stem cell, absorbent cotton and reticulated structure all directly can not contact with cell culture fluid, with reticulated structure, absorbent cotton can also guarantee that perfusion groove support portion does not directly contact with bottom cell perfusion groove 10 inner chamber body the effect playing insulation simultaneously, cell perfusion groove 10 bottom arranges a bracing frame 16, guarantee that cell perfusion groove 10 obtains firm support in certain position aloft, cell perfusion groove 10 two ends connect the second pipe connecting 9 and the 3rd pipe connecting 11 respectively, second pipe connecting 9 the other end is connected with the first peristaltic pump 8, 3rd pipe connecting 11 the other end is connected with the second peristaltic pump 12, waste liquid bottle 15 bottleneck arranges output tube 13 and the second filter 14, second filter 14 can make the pressure in waste liquid bottle 15 identical with the external world, and the bacterial contamination that can completely cut off in outside air, output tube 13 the other end connects the second peristaltic pump 12, one Tissue Culture Plate is set bottom feeder layer cells incubator 5, cell perfusion groove 10 two ends arrange the semi-permeable membranes preventing culturing cell from passing, but semi-permeable membranes can make liquid freely pass through, one annular transparent hole is set in the middle part of cell perfusion groove 10, facilitates observation of cell perfusion groove inner cell to breed situation.
The present invention in use, load in perfusion bottle 1 and be suitable for the nutriment that stem cell cultivates breeding, the element that supplements the nutrients is loaded in feeder layer cells incubator 5, by the effect of the first peristaltic pump 8 and the second peristaltic pump 12, the nutriment in perfusion bottle 1 is flowed in feeder layer cells incubator 5 and cell perfusion groove 10, for stem cell provides sufficient nutrient, improve the cultivation speed of stem cell, the concrete pore size of the semi-permeable membranes bottom feeder layer cells incubator 5 control stem cell can be stoped to pass through but can realize liquid by, similarly, the concrete pore size of semi-permeable membranes at cell perfusion groove 10 two ends control stem cell can be stoped to pass through but can realize liquid by, this perfusion device can at the first controller 4 and second controller 7 and two peristaltic pumps (8, 12) the cultivation breeding of stem cell is completed under acting in conjunction, conveniently record differing temps, during pH value, in feeder layer cells incubator 5 and cell perfusion groove 10, stem cell cultivates the situation of breeding, avoid the error value producing temperature and pH value because manual observation is measured, exported by cell perfusion groove 10 equipment, the cultivation speed of stem cell can be improved.
Above one embodiment of the present of invention have been described in detail, but described content being only preferred embodiment of the present invention, can not being considered to for limiting practical range of the present invention.All equalizations done according to the present patent application scope change and improve, and all should still belong within patent covering scope of the present invention.
Claims (4)
1. stem cell culture perfusing device, is characterized in that: comprise perfusion bottle (1), feeder layer cells incubator (5), cell perfusion groove (10) and waste liquid bottle (15);
Described perfusion bottle (1) bottle mouth position arranges the first filter (2) and input tube (3), described input tube (3) is arranged the first controller (4) that can show pH value and control temperature, described input tube (3) tail end immerses feeder layer cells incubator (5), the first pipe connecting (6) that the feeder layer cells incubator (5) arranging semi-permeable membranes by substrate connects is arranged second controller (7) and the first peristaltic pump (8) that one can show pH value and control temperature, bottom described cell perfusion groove (10) inner chamber body, the sterile-processed absorbent cotton of a regular shape is set, described absorbent cotton top arranges a reticulated structure, described reticulated structure top arranges the perfusion groove support portion of placing cell, and absorbent cotton and reticulated structure all directly can not contact with cell culture fluid, described cell perfusion groove (10) bottom arranges a bracing frame (16), described cell perfusion groove (10) two ends connect the second pipe connecting (9) and the 3rd pipe connecting (11) respectively, described second pipe connecting (9) the other end is connected with described first peristaltic pump (8), described 3rd pipe connecting (11) the other end is connected with described second peristaltic pump (12), described waste liquid bottle (15) bottleneck arranges output tube (13) and the second filter (14), described output tube (13) the other end connects described second peristaltic pump (12).
2. stem cell culture perfusing device according to claim 1, is characterized in that: described feeder layer cells incubator (5) bottom arranges a Tissue Culture Plate and described semi-permeable membranes is positioned on described Tissue Culture Plate.
3. stem cell culture perfusing device according to claim 1, is characterized in that: described cell perfusion groove (10) two ends arrange semi-permeable membranes.
4. stem cell culture perfusing device according to claim 1, is characterized in that: described cell perfusion groove (10) middle part arranges an annular transparent hole.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410812200.7A CN104403946A (en) | 2014-12-22 | 2014-12-22 | Stem cell culture perfusion device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410812200.7A CN104403946A (en) | 2014-12-22 | 2014-12-22 | Stem cell culture perfusion device |
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| Publication Number | Publication Date |
|---|---|
| CN104403946A true CN104403946A (en) | 2015-03-11 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410812200.7A Pending CN104403946A (en) | 2014-12-22 | 2014-12-22 | Stem cell culture perfusion device |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1435482A (en) * | 2002-01-31 | 2003-08-13 | 赛宇细胞科技股份有限公司 | Cell culture device |
| CN201626960U (en) * | 2010-02-01 | 2010-11-10 | 中国农业科学院北京畜牧兽医研究所 | Perfusion device for stem cell culture |
| CN202124632U (en) * | 2011-07-29 | 2012-01-25 | 天津和泽干细胞科技有限公司 | Stem cell culture perfusing device |
| CN204385206U (en) * | 2014-12-22 | 2015-06-10 | 顺昊细胞生物技术（天津）股份有限公司 | Stem cell culture perfusing device |
-
2014
- 2014-12-22 CN CN201410812200.7A patent/CN104403946A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1435482A (en) * | 2002-01-31 | 2003-08-13 | 赛宇细胞科技股份有限公司 | Cell culture device |
| CN201626960U (en) * | 2010-02-01 | 2010-11-10 | 中国农业科学院北京畜牧兽医研究所 | Perfusion device for stem cell culture |
| CN202124632U (en) * | 2011-07-29 | 2012-01-25 | 天津和泽干细胞科技有限公司 | Stem cell culture perfusing device |
| CN204385206U (en) * | 2014-12-22 | 2015-06-10 | 顺昊细胞生物技术（天津）股份有限公司 | Stem cell culture perfusing device |
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| PB01 | Publication | ||
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Application publication date: 20150311 |