CN102399679B - Device for cell microsphere filamentation preparation based on type I collagen gel - Google Patents
Device for cell microsphere filamentation preparation based on type I collagen gel Download PDFInfo
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- CN102399679B CN102399679B CN 201110377051 CN201110377051A CN102399679B CN 102399679 B CN102399679 B CN 102399679B CN 201110377051 CN201110377051 CN 201110377051 CN 201110377051 A CN201110377051 A CN 201110377051A CN 102399679 B CN102399679 B CN 102399679B
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- 102000012422 Collagen Type I Human genes 0.000 title claims abstract description 37
- 108010022452 Collagen Type I Proteins 0.000 title claims abstract description 37
- 239000000512 collagen gel Substances 0.000 title claims abstract description 23
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 239000004005 microsphere Substances 0.000 title abstract description 10
- 238000007493 shaping process Methods 0.000 claims abstract description 7
- 102000008186 Collagen Human genes 0.000 claims description 31
- 108010035532 Collagen Proteins 0.000 claims description 31
- 229920001436 collagen Polymers 0.000 claims description 31
- 239000000835 fiber Substances 0.000 claims description 31
- 230000002572 peristaltic effect Effects 0.000 claims description 15
- 230000000694 effects Effects 0.000 claims description 8
- 235000015097 nutrients Nutrition 0.000 claims description 5
- 230000015572 biosynthetic process Effects 0.000 claims description 4
- 230000001105 regulatory effect Effects 0.000 claims description 3
- 238000007789 sealing Methods 0.000 claims description 3
- 230000007306 turnover Effects 0.000 claims description 3
- 230000005514 two-phase flow Effects 0.000 claims description 3
- 238000000034 method Methods 0.000 abstract description 7
- 238000011081 inoculation Methods 0.000 abstract description 5
- 239000003814 drug Substances 0.000 abstract description 4
- 230000001172 regenerating effect Effects 0.000 abstract description 4
- 210000000056 organ Anatomy 0.000 abstract description 3
- 230000003321 amplification Effects 0.000 abstract description 2
- 239000012620 biological material Substances 0.000 abstract description 2
- 239000006285 cell suspension Substances 0.000 abstract description 2
- 238000003199 nucleic acid amplification method Methods 0.000 abstract description 2
- 238000010276 construction Methods 0.000 abstract 1
- 230000007547 defect Effects 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 116
- 238000005516 engineering process Methods 0.000 description 3
- 238000002659 cell therapy Methods 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000010041 electrostatic spinning Methods 0.000 description 1
- 235000015110 jellies Nutrition 0.000 description 1
- 239000008274 jelly Substances 0.000 description 1
- 238000002074 melt spinning Methods 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 239000002861 polymer material Substances 0.000 description 1
- 229920002635 polyurethane Polymers 0.000 description 1
- 239000004814 polyurethane Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000009987 spinning Methods 0.000 description 1
- 230000003019 stabilising effect Effects 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- 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
- C12M41/36—Means for regulation, monitoring, measurement or control, e.g. flow regulation of concentration of biomass, e.g. colony counters or by turbidity measurements
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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
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/16—Particles; Beads; Granular material; Encapsulation
- C12M25/20—Fluidized bed
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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
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
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- General Health & Medical Sciences (AREA)
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Abstract
The invention relates to a device for cell microsphere filamentation preparation based on type I collagen gel. In the traditional method, the large-scale amplification of cells is carried out through passage, manually harvested seed cells are also compounded with biomaterials through manual inoculation, the operation is simple and complicated, the pollution is easy to cause, and the cell inoculation is not uniform. The device comprises a cell microsphere filamentation device, a control unit and a measuring unit, wherein the cell microsphere filamentation device consists of three parts: a cell microsphere and type I collagen gel mixing positioning mechanism, a cell microsphere silk thread shaping mechanism and a cell microsphere silk thread collecting mechanism. The device has the advantages that cell microsphere active silk threads covered by type I collagen can be prepared, the defect that cell suspension can not control the cell position state in the cell inoculation and the cell treatment is overcome, a large number of different kinds of cell microsphere single silks are provided for the precise construction of complicated tissues and organs, and the device is provided for tissue engineering and the clinical application of regenerative medicine.
Description
Technical field
The invention belongs to biomedical engineering field, relate to the device that a kind of cell microsphere becomes the silk preparation outward.Particularly relate to a kind of for the device with the active silk thread of the coated type i collagen gel formation of cell microballoon.Be suitable for the cell microballoon silk thread that the external preparation of biomedical engineering field has cytoactive, be applied to the accurate inoculation of tissue engineering complex tissue and organ and the cell therapy of regenerative medicine.
Background technology
In the clinical application of organizational project and regenerative medicine, traditional method is to carry out the extensive amplification of cell by going down to posterity, the seed cell of manual results is also inoculate and biomaterial carries out compoundly by craft, and loaded down with trivial details, easy pollution simple to operate, cell are inoculated inhomogeneous.In addition, at present the one-tenth silk method of polymer material polyurethane is solvent spinning and melt-spinning, and these two kinds of methods are owing to will carry out in high temperature and solution, and cell can not be survived and can not be prepared cell microballoon silk thread; Electrostatic spinning technique is the spining technology that carries out on nanoscale, because the diameter of silk thread is also thinner than cell, can not carry out the preparation that the cell microballoon becomes silk.Can't satisfy the demand that organizational project builds the tissue of complex shape.
Summary of the invention
The purpose of this invention is to provide a kind of device that the coated type i collagen gel of cell microballoon is prepared the active silk thread of cell microballoon, can obtain to carry out position control to the cell microballoon based on the active monofilament of the cell microballoon of type i collagen gel.For cell microballoon filament forming device, can control flow state and the flow velocity of cell microballoon and type i collagen gel, control the temperature that the cell microballoon becomes the silk process, form the cell monofilament with the jelly shape of cytoactive of different diameter.
The technical scheme that technical solution problem of the present invention is taked is:
Become the device of silk preparation based on type i collagen gel cell microballoon, comprise cell microballoon filament forming device, control unit and measuring unit.
Described cell microballoon filament forming device is comprised of cell microballoon and type i collagen gel mixed positioning mechanism, cell microballoon silk thread shaping mechanism and cell microballoon silk thread collecting mechanism three parts.
Described cell microballoon comprises that with type i collagen gel mixed positioning mechanism right peristaltic pump, mixing vessel upper cover, type i collagen-nutrient solution right entry, cell microballoon become a silk platform, cell microballoon-type i collagen to become fiber tube, left peristaltic pump, cell microballoon entrance, mixing vessel, type i collagen-nutrient solution left entry.
cell microballoon entrance is positioned at the top of mixing vessel upper cover, mixing vessel upper cover and mixing vessel are connected to form the gnotobasis of a sealing, type i collagen-nutrient solution right entry and type i collagen-nutrient solution left entry lay respectively at tangent position, mixing vessel side with mixing vessel to being communicated with, make the type i collagen that enters mixing vessel-nutrient solution form the state of helicoidal flow, type i collagen-nutrient solution right entry is connected with left peristaltic pump with right peristaltic pump respectively with the entrance of type i collagen-nutrient solution left entry, can control the speed of eddy flow by the flow of regulating two peristaltic pumps, the cell microballoon becomes fiber tube through flowing and entering cell microballoon-type i collagen under cell microballoon entrance inflow mixing vessel and helicoidal flow junction formation screw, the cell microballoon must be in flow velocity the fastest central position.By cell microballoon and the two-phase flow of type i collagen in cell microballoon-type i collagen becomes fiber tube, can form the structure of the coated cell microballoon silk thread of type i collagen, so just can locate the cell microballoon.Whole cell microballoon is positioned at the cell microballoon with type i collagen gel mixed positioning mechanism and becomes on the silk platform.
Described cell microballoon silk thread shaping mechanism comprises that the cell microballoon becomes silk platform, 37 ℃ of constant temperature uncontaminated air right entries, 37 ℃ of constant temperature uncontaminated air left entries, supporting bar, cell microballoon-type i collagens to become fiber tube, 37 ℃ of uncontaminated air typing chambers and 37 ℃ of constant temperature uncontaminated air outlets.
37 ℃ of uncontaminated airs are finalized the design, and the chamber is positioned at cell microballoon one-tenth silk platform below and supporting bar is inner, both sides respectively with 37 ℃ of constant temperature uncontaminated air right entries with are connected a ℃ constant temperature uncontaminated air left entry and are connected, cell microballoon-type i collagen becomes fiber tube to pass 37 ℃ of uncontaminated airs typing 37 ℃ of chambers constant temperature uncontaminated air outlets to extend out.Like this, 37 ℃ of uncontaminated airs enter 37 ℃ of uncontaminated airs typing chambers by two side entrances, the cell microballoon silk thread of type i collagen gel pack quilt is formed the effect of typing, and, 37 ℃ of uncontaminated airs flow out through 37 ℃ of constant temperature uncontaminated air outlets, also can play the effect of further typing and location to become the cell microballoon silk thread that fiber tube flows out from cell microballoon-type i collagen.
Described cell microballoon silk thread collecting mechanism comprises that base, cell microballoon-type i collagen become fiber tube and cell microballoon silk thread collection container.
Cell microballoon silk thread collection container is positioned at the top of base, cell microballoon-type i collagen becomes fiber tube to be positioned at again the top of cell microballoon silk thread collection container, when cell microballoon silk thread becomes when flowing out fiber tube from cell microballoon-type i collagen, can enter in the cell microballoon silk thread collection container that contains nutrient solution and collect.
Described control unit comprises touch-screen, programmable logic controller, motor and speed regulator; Can be to the turnover parameter setting of cell microballoon filament forming device and operation by touch-screen, the instruction of touch-screen is passed to programmable logic controller by data line, programmable logic controller is completed control to motor according to software prepared in advance, and speed regulator can be carried out speed governing to motor.
Described measuring unit comprises pressure transmitter, flow sensor, temperature sensor, data collecting card, Survey Software; The data of pressure transmitter and flow sensor collection pass to data collecting card by data line, and data collecting card by Survey Software with data presentation on computers.
Patent of the present invention is compared with background technology, the beneficial effect that has is: can prepare the active silk thread of the coated cell microballoon of type i collagen, overcome the state of cell suspension uncontrollable cell position in cell inoculation and cell therapy, provide a large amount of different types of cell microballoon monofilament for accurately building complex organization and organ, installed preferably for the clinical application of organizational project and regenerative medicine provides a kind of.
Description of drawings
Fig. 1 is that the present invention is about cell microballoon filament forming device shape assumption diagram.
Embodiment
The invention will be further described below in conjunction with accompanying drawing.
As shown in Figure 1, become the device of silk preparation based on type i collagen gel cell microballoon, comprise cell microballoon filament forming device, control unit and three parts of measuring unit.
Cell microballoon filament forming device comprises: cell microballoon and type i collagen gel mixed positioning mechanism, cell microballoon silk thread shaping mechanism and cell microballoon silk thread collecting mechanism three parts form.
The cell microballoon comprises with type i collagen gel mixed positioning mechanism: right peristaltic pump 1, mixing vessel upper cover 2, type i collagen-nutrient solution right entry 3, cell microballoon become a silk platform 4, cell microballoon-type i collagen to become fiber tube 7, left peristaltic pump 9, cell microballoon entrance 10, mixing vessel 11, type i collagen-parts such as nutrient solution left entry 12 to consist of.cell microballoon entrance 10 is positioned at the top of mixing vessel upper cover 2, mixing vessel upper cover 2 and mixing vessel 11 are connected to form the gnotobasis of a sealing, type i collagen-nutrient solution right entry 3 and type i collagen-nutrient solution left entry 12 lay respectively at tangent position, mixing vessel 11 sides with mixing vessel 11 to being communicated with, make the type i collagen that enters mixing vessel 11-nutrient solution form the state of helicoidal flow, type i collagen-nutrient solution right entry 3 is connected with left peristaltic pump 9 with right peristaltic pump 1 respectively with the entrance of type i collagen-nutrient solution left entry 12, can control the speed of eddy flow by the flow of regulating two peristaltic pumps, the cell microballoon becomes fiber tube 7 through flowing and entering cell microballoon-type i collagen under cell microballoon entrance 10 inflow mixing vessels 11 and helicoidal flow junction formation screw, to be parabolical shape due to stabilising liq at the flow condition of common pipe, the flow velocity at pipe center is the fastest, when the cell microballoon enters cell microballoon-type i collagen and becomes in fiber tube 7, the cell microballoon must be in flow velocity the fastest central position.And the helical flow kinetic energy of outside type i collagen-nutrient solution is with the even coated cell microballoon of type i collagen, become two-phase flow in fiber tube 7 by cell microballoon and type i collagen at cell microballoon-type i collagen, can form the structure of the coated cell microballoon silk thread of type i collagen, so just can locate the cell microballoon.Whole cell microballoon is positioned at the cell microballoon with type i collagen gel mixed positioning mechanism and becomes on silk platform 4.
Cell microballoon silk thread shaping mechanism comprises: the cell microballoon becomes a silk platform 4,37 ℃ of constant temperature uncontaminated air right entries 5,37 ℃ of constant temperature uncontaminated air left entries 14, supporting bar 6, cell microballoon-type i collagens to become fiber tube 7,37 ℃ of uncontaminated airs typing chambers 13,37 ℃ of parts such as constant temperature uncontaminated air outlet 15 to consist of.
37 ℃ of uncontaminated airs typing chamber 13 is positioned at the cell microballoon and becomes silk platform 4 belows and supporting bar 6 inside, both sides respectively with 37 ℃ of constant temperature uncontaminated air right entries 5 with are connected a ℃ constant temperature uncontaminated air left entry 14 and are connected, cell microballoon-type i collagen becomes fiber tube 7 to pass 37 ℃ of uncontaminated airs typing chambers 13 to extend out through 37 ℃ of constant temperature uncontaminated airs outlets 15.Like this, 37 ℃ of uncontaminated airs enter 37 ℃ of uncontaminated airs typing chambers 13 by two side entrances, the cell microballoon silk thread of type i collagen gel pack quilt is formed the effect of typing, and, 37 ℃ of uncontaminated airs flow out through 37 ℃ of constant temperature uncontaminated air outlets 15, also can play the effect of further typing and location to become the cell microballoon silk thread that fiber tube 7 flows out from cell microballoon-type i collagen.
Cell microballoon silk thread collecting mechanism comprises that base 8, cell microballoon-type i collagen become the parts such as fiber tube 7, cell microballoon silk thread collection container 16 to form.
Cell microballoon silk thread collection container 16 is positioned at the top of base 8, cell microballoon-type i collagen becomes fiber tube 7 to be positioned at again the top of cell microballoon silk thread collection container 16, when cell microballoon silk thread becomes when flowing out fiber tube 7 from cell microballoon-type i collagen, can enter in the cell microballoon silk thread collection container 16 that contains nutrient solution and collect.
Control unit is comprised of elements such as touch-screen, programmable logic controller, motor, speed regulator.Can be to cell microballoon filament forming device turnover parameter setting and operation by touch-screen, the instruction of touch-screen is passed to controller by data line, and controller is completed control to motor according to software prepared in advance, and speed regulator can be carried out speed governing to motor.
Measuring unit partly is comprised of pressure transmitter, flow sensor, data collecting card, Survey Software etc.The data of pressure transmitter and flow sensor collection pass to data collecting card by data line, and data collecting card by Survey Software with data presentation on computers.
The working process of this cell microballoon filament forming device is as follows:
1, cultured cell microballoon is injected through cell microballoon entrance 10, simultaneously, the type i collagen gelating soln is injected mixing vessel 11 by type i collagen-nutrient solution right entry 3 and type i collagen-nutrient solution left entry 12, form flowing under screw, and become fiber tube 7 with cell microballoon junction inflow cell microballoon-type i collagen, owing to having formed time spiral motion, the cell microballoon can be fixed on middle position and evenly moving downward that cell microballoon-type i collagen becomes fiber tube 7 and enter 37 ℃ of uncontaminated airs typing chambers 13.
2,37 ℃ of constant temperature uncontaminated airs are flowed into 37 ℃ of uncontaminated airs typing chambers 13 through 37 ℃ of constant temperature uncontaminated air right entries 5 and 37 ℃ of constant temperature uncontaminated air left entries 14, at last, flow out straight down by 37 ℃ of constant temperature uncontaminated air outlets 15, to the effect that whole cell microballoon-type i collagen becomes fiber tube 7 to be incubated, make the active silk thread typing of the coated cell microballoon of type i collagen flow out.
3, the active silk thread of the cell microballoon that flows out enters the cell microballoon silk thread collection container 16 of filling nutrient solution, the silk thread that has obtained to have cell microballoon activity.
Claims (1)
1. become the device of silk preparation based on type i collagen gel cell microballoon, comprise cell microballoon filament forming device, control unit and measuring unit, it is characterized in that:
Described cell microballoon filament forming device is comprised of cell microballoon and type i collagen gel mixed positioning mechanism, cell microballoon silk thread shaping mechanism and cell microballoon silk thread collecting mechanism three parts;
Described cell microballoon comprises that with type i collagen gel mixed positioning mechanism right peristaltic pump (1), mixing vessel upper cover (2), type i collagen-nutrient solution right entry (3), cell microballoon become a silk platform (4), cell microballoon-type i collagen to become fiber tube (7), left peristaltic pump (9), cell microballoon entrance (10), mixing vessel (11), type i collagen-nutrient solution left entry (12);
cell microballoon entrance (10) is positioned at the top of mixing vessel upper cover (2), mixing vessel upper cover (2) and mixing vessel (11) are connected to form the gnotobasis of a sealing, type i collagen-nutrient solution right entry (3) and type i collagen-nutrient solution left entry (12) lay respectively at tangent position, mixing vessel (11) side and are connected with mixing vessel (11), make the type i collagen that enters mixing vessel (11)-nutrient solution form the state of helicoidal flow, type i collagen-nutrient solution right entry (3) is connected with left peristaltic pump (9) with right peristaltic pump (1) respectively with the entrance of type i collagen-nutrient solution left entry (12), can control the speed of eddy flow by the flow of regulating two peristaltic pumps, the cell microballoon becomes fiber tube (7) through flowing and entering cell microballoon-type i collagen under cell microballoon entrance (10) inflow mixing vessel (11) and helicoidal flow junction formation screw, the cell microballoon must be in flow velocity the fastest central position, become two-phase flow in fiber tube (7) by cell microballoon and type i collagen at cell microballoon-type i collagen, can form the structure of the coated cell microballoon silk thread of type i collagen, so just can locate the cell microballoon, whole cell microballoon is positioned at the cell microballoon with type i collagen gel mixed positioning mechanism and becomes on silk platform (4),
Described cell microballoon silk thread shaping mechanism comprises that the cell microballoon becomes silk platform (4), 37 ℃ of constant temperature uncontaminated air right entries (5), 37 ℃ of constant temperature uncontaminated air left entries (14), supporting bar (6), cell microballoon-type i collagen to become fiber tube (7), 37 ℃ of uncontaminated air typing chambers (13) and 37 ℃ of constant temperature uncontaminated airs to export (15);
37 ℃ of uncontaminated airs typing chambers (13) are positioned at the cell microballoon and become silk platform (4) below and supporting bar (6) inside, both sides respectively with 37 ℃ of constant temperature uncontaminated air right entries (5) with are connected a ℃ constant temperature uncontaminated air left entry (14) and are connected, cell microballoon-type i collagen becomes fiber tube (7) to pass 37 ℃ of uncontaminated airs typing chambers (13) to export (15) through 37 ℃ of constant temperature uncontaminated airs and extend out; Like this, 37 ℃ of uncontaminated airs enter 37 ℃ of uncontaminated airs typing chambers (13) by two side entrances, the cell microballoon silk thread of type i collagen gel pack quilt is formed the effect of typing, and, 37 ℃ of uncontaminated airs export (15) through 37 ℃ of constant temperature uncontaminated airs and flow out, and also can play the effect of further typing and location to become the cell microballoon silk thread that fiber tube (7) flows out from cell microballoon-type i collagen;
Described cell microballoon silk thread collecting mechanism comprises that base (8), cell microballoon-type i collagen become fiber tube (7) and cell microballoon silk thread collection container (16);
Cell microballoon silk thread collection container (16) is positioned at the top of base (8), cell microballoon-type i collagen becomes fiber tube (7) to be positioned at again the top of cell microballoon silk thread collection container (16), become from cell microballoon-type i collagen when cell microballoon silk thread when flowing out fiber tube (7), can enter in the cell microballoon silk thread collection container (16) that contains nutrient solution and collect;
Described control unit comprises touch-screen, programmable logic controller, motor and speed regulator; Can be to the turnover parameter setting of cell microballoon filament forming device and operation by touch-screen, the instruction of touch-screen is passed to programmable logic controller by data line, programmable logic controller is completed control to motor according to software prepared in advance, and speed regulator can be carried out speed governing to motor;
Described measuring unit comprises that the data of pressure transmitter, flow sensor, temperature sensor, data collecting card, Survey Software, pressure transmitter and flow sensor collection pass to data collecting card by data line, and data collecting card by Survey Software with data presentation on computers.
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| Application Number | Priority Date | Filing Date | Title |
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| CN 201110377051 CN102399679B (en) | 2011-11-24 | 2011-11-24 | Device for cell microsphere filamentation preparation based on type I collagen gel |
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| CN 201110377051 CN102399679B (en) | 2011-11-24 | 2011-11-24 | Device for cell microsphere filamentation preparation based on type I collagen gel |
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| CN102399679B true CN102399679B (en) | 2013-05-22 |
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| CN102853939A (en) * | 2012-09-03 | 2013-01-02 | 江苏大学 | Real-time monitoring device and method of vinegar grains fermentation process |
| CN105726313A (en) * | 2016-04-25 | 2016-07-06 | 上海东富龙科技股份有限公司 | Integrated microsphere preparation device |
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Effective date of registration: 20161012 Address after: 411101 high tech building, Furong Middle Road, high tech Zone, Hunan, Xiangtan, China, No. 9 Patentee after: Xiangtan poly energy saving environmental technology Co.,Ltd. Address before: Hangzhou City, Zhejiang province 310018 Xiasha Higher Education Park No. 2 street Patentee before: HANGZHOU DIANZI University |
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