CN104232484A - Cell co-culture model and preparation method - Google Patents
Cell co-culture model and preparation method Download PDFInfo
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Abstract
The invention discloses a cell co-culture model which comprises a matrix, wherein a fiber culture is constructed in the matrix, two ends of the fiber culture extend out of the outer wall of the matrix, and the fiber culture is made of a gel material; two or more channels for cell co-culturing are arranged in the fiber culture, and are independent from each other, and each channel is provided with an inlet and an outlet which are positioned in two ends of the fiber culture; the gel material is a crosslinked product of sodium alginate and calcium chloride. The invention further discloses a preparation method of the model. The cell co-culture model can establish two and more than two cell non-contact type co-culture systems, the true growth status of cells in the body can be simulated to the greatest extent, the dynamic co-culture of the cells can be realized, and the observation and subsequent detection and analysis are facilitated. In addition, the preparation method of the cell co-culture model is simple to operate, and high in production efficiency by utilizing an island-shaped spray head-based printing device.
Description
Technical field
The invention belongs to Cell Co culturing Techenique field, especially relate to a kind of cell co-culture model and preparation method.
Background technology
Cell in vitro Dual culture is by the technology of two or more cell co-culture in same environment.The microenvironment of Cell Co culturing Techenique energy analogue body inner cell normal activity; be conducive to the interaction between observation of cell and culture environment, between cell and cell; because it has the advantage of antimer environment better, be in this way widely used in INVENTIONModern cell research.
The method of current co-culture of cells mainly contains direct contact type Dual culture and non-direct contact type Dual culture.Direct contact type Dual culture is mixed by dissimilar cell to carry out co-cultivation in same culture system, and two kinds of cells are directly contacted, and produced the cells produce factor etc. interacted by paracrine, Autocrine.Its shortcoming is that directly contact occurs dissimilar cell, thoroughly can not separate, be unfavorable for that subsequent analysis is observed.Non-direct contact type Dual culture is seeded on different carriers respectively by two or more cell, then these carriers are placed in same culture environment, in co-culture system, the impact of one on another person is interacted by the cytokine of paracrine, but both do not contact.Because two kinds of cells are easily separated, be convenient to observe and do not affect follow-up detection.
Contactless cell co-culture method conventional at present uses Transwell cell Dual culture method: based on the cup-formed device having permeability, cup bottom puts the film that has permeability, Transwell cell is put into culture plate, cell A kind is in upper indoor, lower floor repopulating cell B, composition in the nutrient solution of cell B can have influence on the cell of indoor, thus can study the impact of the materials onto cells A that cell B secretes or metabolism produces.Vertical stratification in this method cannot be got rid of gravity interference for the detection of cell movement and also be unfavorable for carrying out detection of dynamic, and expensive.
Because existing cell co-culture method exists certain defect, limit co-culture of cells progress of research, the contactless cell co-culture method of therefore cheap, the convenient practicality of cost of development is following important development direction.
Summary of the invention
In order to overcome the technical problem existed in prior art, the invention provides a kind of cell co-culture model, by this cell co-culture model, can ensure that different cell does not directly contact, and observation and the analysis of the interaction behavior between cell and cell can be realized, reduce the impact of the extraneous factors such as gravity.
The present invention additionally provides a kind of preparation method of cell co-culture model simultaneously, and the method can produce practical and controlled contactless cell co-culture model, and production efficiency is high, economical convenient.
A kind of cell co-culture model, comprises matrix, and be built with the fiber cultivation body that matrix outer wall is stretched out at two ends in described matrix, this fiber is cultivated body and is made up of gel material; Described fiber is cultivated in body and is provided with two or more passages for co-culture of cells, separate between each passage, and each passage has and lays respectively at the entrance and exit that body two ends cultivated by fiber; Described gel material is the cross-linking products of sodium alginate and calcium chloride.
Cell co-culture model of the present invention adopts the fiber manufactured by bio-compatibility gelatinous material to cultivate body as the carrier of cell, by cell seeding in adjacent passage, utilizes the permeability of gel, dissimilar cellular compartments is driven into row and cultivates.Because hydrogel has permeability, molecule can exchange through gel partition, does not need to offer separately to exchange duct.Nutrient solution containing target cell in passage, utilize the adherent property of cell, the cell attachment of Dual culture is on the inwall of passage.Inventive gel material is the cross-linking products of sodium alginate and calcium chloride, and the fiber cultivation body solid section with permeability is alginate calcium, and the structural strength of formation is higher.
Described matrix is formed by gel material or is made up of polydimethylsiloxane.Described gelatinous material can adopt the cross-linking products of sodium alginate and calcium chloride.This model manufactures the matrix of coated tubular fibre structure by the gelatinous material of bio-compatibility, utilizes matrix to have certain intensity, is structured in by co-culture model in a stable environment.
Body cultivated by described fiber is that roundabout shape is arranged.Fiber is cultivated body and is adopted in roundabout shape layout, further reduces the volume of whole model, meets the demand of various occasion simultaneously.
A kind of preparation method of cell co-culture model, the method utilizes the printing technique based on island shower nozzle to manufacture containing multichannel culture model, then in adjacency channel, plant different types of cell respectively, realize contactless co-culture of cells, the method is specifically carried out as follows:
(1) configure sodium alginate soln I and calcium chloride solution I, configure in sodium alginate soln I and calcium chloride solution I process, magnetic stirrer can be adopted to stir;
(2) sodium alginate soln I and calcium chloride solution I is passed into respectively in syringe corresponding to the different passages of island printing head;
Described island printing head comprises: two or more endoporus, and quantity and the structure of the quantity of endoporus and structure and passage match; Be enclosed in the exit orifice of endoporus periphery; The interior passageway be communicated with respectively with endoporus; The exterior passage way be communicated with exit orifice; Described interior passageway and exterior passage way are connected with respective syringe respectively, fill calcium chloride solution I in the syringe be wherein communicated with interior passageway, fill sodium alginate soln I in the syringe be communicated with exterior passage way;
(3) body material is configured;
If Choice of substrate materials sodium alginate soln and calcium chloride solution cross-linking products, then layoutprocedure is as follows:
Configure another and organize certain density sodium alginate soln II, utilize magnetic stirrer to stir;
If polydimethylsiloxane (PDMS) selected by body material, then layoutprocedure is as follows: mixed according to certain ratio with solidifying agent supporting with it in prior art by polydimethylsiloxane, the mass ratio of polydimethylsiloxane and solidifying agent is 8-15:1, as preferably, mass ratio is 10:1.
(4) body material that step (3) configures is poured in the container of motion platform, ensure that body material is evenly distributed in a reservoir and has smooth surface simultaneously, form the first layer base material layer; In this step, Horizontal vibration device can be utilized sodium alginate to be evenly distributed in a reservoir and to have smooth surface;
(5) utilize the printing head in step (2) to print fiber according to desired trajectory on the first layer base material layer and cultivate body entity, wherein fiber is cultivated body entity periphery and is printed sodium alginate soln I, and fiber cultivates body entity house print for the formation of two or more calcium chloride solution I for the passage of co-culture of cells; In this step print procedure, moving movement platform can be utilized, the movement velocity of adjustment movement platform, make many passages of formation be deposited on the surface of the first layer base material layer in step (4) with certain track;
(6) body material that step (3) configures is continued to pour in container, cover the fiber formed and cultivate on body entity; In this step, Horizontal vibration device can be utilized second layer base material layer to be evenly distributed in a reservoir and to have smooth surface;
(7) process is cured to the chip first product with fiber cultivation body structure that step (6) obtains, obtains many cells co-culture model.
Solidification treatment can be selected respectively according to body material difference:
When Choice of substrate materials sodium alginate soln and calcium chloride solution cross-linking products, configurablely the calcium chloride solution II of crosslinked action can be there is with sodium alginate soln II; Body material solidification process can be realized by spraying calcium chloride solution II; In this solidification process, calcium chloride solution II is first sprayed on the fiber that has that (6) obtain and cultivates on the chip first product of body structure by general employing, makes the sodium alginate cross-linking on surface become gel, realizes Procuring; Then the chip first product of whole Procuring is immersed in the calcium chloride solution II configured, make total be cross-linked into the gel structure with some strength completely, finally obtain the Gel chips containing multiple passage;
As Choice of substrate materials polydimethylsiloxane (PDMS), curing is as follows: the chip cultivating body structure containing fiber is put into baking oven, and temperature is set to 65-85 DEG C, and set time is 60-80 minute.
The concentration of the sodium alginate soln I in step (1) depends on intensity and and the porosity of multiple passages of required preparation.The concentration of sodium alginate soln I is too low, and the intensity of multiple passages of formation is too little, and gel porosity is too large; The concentration of sodium alginate soln I is too high, and the composite hollow gel-strength of formation is too large, and gel porosity is too little, and sodium alginate viscosity is too large, does not utilize shower nozzle to spray.As preferably, utilize deionized water as solvent, the mass percent concentration of configuration sodium alginate soln I is 1%-3%, the moderate strength of many passages of formation like this and porosity meets permeability requirement.
The concentration of the calcium chloride solution I in step (1) depends on external diameter and the wall thickness of many passages of required preparation.The concentration of calcium chloride solution I is too low, and external diameter and the wall thickness of multiple passages of formation are too little; The concentration of calcium chloride solution I is too high, and external diameter and the wall thickness of multiple passages of formation are too large.As preferably, utilize deionized water as solvent, the mass percent concentration of configuration calcium chloride solution I is 2%-5%, and external diameter and the wall thickness of the multiple passages formed like this are moderate, are conducive to cell and exchange with intercellular.
The syringe used in step (2) is generally connected with syringe pump, and syringe pump is the autonomous triple channel syringe pump manufactured, and can control the flow velocity of each passage respectively.Utilize the convert rotational motion of three reducing motors to be the uniform motion that the translational motion of three leading screws realizes syringe, it is adjustable that the rotating speed controlling motor respectively by corresponding sequence of control realizes speed.The syringe containing sodium alginate soln be connected with shower nozzle exit orifice, to be all placed on this triple channel syringe pump with multiple syringes of island shower nozzle endoporus.
In step (3), another group sodium alginate soln II and calcium chloride solution II is gel-in-matrix in order to form coated multiple passage, so relative concentration more greatly, to meet requirement of strength.As preferably, utilize deionized water as solvent, the mass percent concentration of configuration sodium alginate soln II is 3%-5%, and the mass percent concentration of configuration calcium chloride solution II is 5%-8%.
When body material adopts the cross-linking products of sodium alginate and calcium chloride, in step (4) and step (6), can rational coated multiple passage and will be easy to follow-up being cross-linked for the height of two-layer sodium alginate.As preferably, the height of two-layer sodium alginate is respectively 3-5mm.
The principle obtaining multiple passage in step (5) is: the calcium chloride solution that the sodium alginate soln flowed out by shower nozzle exit orifice flows out with shower nozzle endoporus contacts, first there is crosslinking reaction in surface, form one deck calcium alginate gel, this layer of calcium alginate gel makes sodium alginate and calcium chloride solution be separated in both sides, because the hole of the gel coat formed is very little, calcium ion can only be allowed to pass through, Lalgine ion is not allowed to pass through, so the sodium alginate that the calcium ion of internal layer passes gel coat and outside continues to react, until calcium ion complete reaction, final formation outer solid part is calcium alginate gel, internal void is divided into the tubular fibre structure of the aqueous solution.
The velocity of flow of the liquid be connected with shower nozzle exit orifice, endoporus controlled by syringe pump in step (5) will meet formed tubular fibre structure and be cross-linked evenly, and intensity is better.The most preferred, the flow velocity of the sodium alginate soln I be connected with shower nozzle exit orifice is 0.5-2ml/min, and the calcium chloride solution flow velocity be connected with multiple shower nozzle endoporus keeps identical, is 1-5ml/min.
In step (5), the speed of motion platform will be mated with the flow velocity of solution, to form uniform straight line.As preferably, the speed of motion platform is 1000-1600mm/min.
The movement locus that the fiber cultivation body formed in step (5) is deposited on sodium alginate surface in step (5) can have any shape.The most preferred, movement locus is S shape.
For two passage culture models, during the actual use of cell co-culture model of the present invention:
I nutrient solution containing cell A and cell B is expelled to fiber and cultivates in the adjacent passage of two of body by () respectively, just obtain cell A and cell B co-culture model after leaving standstill for some time;
(ii) cell culture fluid is poured into continuously fiber to cultivate in the passage of body, realize the dynamic Dual culture of cell.
In step (i), repopulating cell A and cell B in the adjacent passage of two of body cultivated by fiber, can contain the nutrient solution of cell A and cell B by direct injector to inject, also can use ejection of syringe pump.As preferably, select the mode repopulating cell of syringe pump, only need use two passages in the triple channel syringe pump in the present invention, flow velocity is 2-5ml/min, and this like cell is evenly distributed in passage, is easy to control.
In step (i), the object leaving standstill for some time is to make cell completely adherent.As preferably, time of repose is 10-16 hour.
In step (ii), cell culture fluid is poured into continuously fiber to cultivate in the passage of body, acyclic and circulation two kinds of logical liquid modes can be adopted.As preferably, selecting the logical liquid mode of circulation, pour into cell culture fluid in cultivate body to fiber two passages with utilizing dual-channel peristaltic pump continuous circulation, like this, the secretory product in cell cultivation process can not run off, and forms a stable culture environment.
Adopt method of the present invention, if two or more cell co-culture models will be obtained, only need increase the quantity of shower nozzle endoporus and corresponding feed flow and control device.
Cell co-culture model of the present invention compared with prior art, has the following advantages:
(1) the present invention can set up two kinds and the contactless co-culture system of two or more cells.
(2) co-culture system of the present invention is multicell parallel construction, and can observe the situation of two kinds of cells under inverted microscope, the cell simultaneously can collecting multicell quantitatively learns the detection of index.
(3) cell culture fluid can pour in the passage of fiber cultivation body by the present invention continuously, can realize the dynamic Dual culture of cell.
(4) cell co-culture model manufacture craft of the present invention is simple, and production efficiency is high.
(5) the present invention utilizes bio-compatibility gelatinous material as cell and intercellular barrier film, and material obtains easily, avoids using expensive material and technique.
Accompanying drawing explanation
Fig. 1 is the structural representation of a kind of cell co-culture model of the present invention.
Fig. 2 is the cross sectional plan view of cell co-culture model shown in Fig. 1.
The A-A cross-sectional schematic that Fig. 3 is cell co-culture model shown in Fig. 2.
Fig. 4 is the structural representation of cell co-culture model using state of the present invention.
Fig. 5 is the sectional structure chart of cell co-culture model of the present invention for three kinds of co-culture of cells.
Fig. 6 is the type printer structural representation wanted required for the present invention.
Fig. 7 is the structural representation of the island shower nozzle wanted required for the present invention.
Fig. 8 is the shower nozzle of island shown in Fig. 7 B-B cross-sectional schematic.
In above-mentioned accompanying drawing: 1 is fiber cultivation body, 2 is that covered fiber cultivates the gel-in-matrix of body, 3 is scleroblast, 4 is osteoclast, and 5 for holding the container for storing liquid containing osteoblasts cultivation liquid, and 6 for holding the container for storing liquid of Osteoclast culture liquid, 7 is dual-channel peristaltic pump, 8 is cell C, and 9 is island shower nozzle, and 10 is triple channel syringe pump, 11 is motion platform, 12 is the exit orifice of island shower nozzle, and 13 is the endoporus of island shower nozzle, and 14 is the endoporus of island shower nozzle, 15 is interior passageway, 16 is exterior passage way, and 17 is cell A, and 18 is cell B.
Embodiment
Embodiment 1
The technical scheme concrete based on the cell co-culture model of Gel chips as of the present invention in Fig. 1-Fig. 3 is as follows:
A kind of cell co-culture model, comprises gel-in-matrix 2, and be built with the fiber cultivation body 1 that gel-in-matrix 2 outer wall is stretched out at two ends in gel-in-matrix 2, body 1 cultivated by fiber is tubular fibre structure, and this fiber is cultivated body 1 and is made up of gel material; Fiber is cultivated in body 1 and is provided with two passage 1a for co-culture of cells, separate between each passage 1a, and each passage 1a has the entrance 1b and outlet 1c that lay respectively at fiber cultivation body 1 two ends; Gel material is the cross-linking products of sodium alginate and calcium chloride; Passage 1a is that roundabout shape is arranged in gel-in-matrix 2.
The tubular fibre structure that this model is manufactured by bio-compatibility gelatinous material, as the carrier of cell, by cell seeding in adjacent hollow gelled fibre passage 1a, utilizes the permeability of gel, dissimilar cellular compartments is driven into row and cultivates; This model manufactures the gel-in-matrix of coated tubular fibre structure by biocompatible material, utilizes gel-in-matrix to have certain intensity, is structured in by co-culture model in a stable environment.
The solid section of fiber cultivation body 1 is the hydrogel with some strength after being cross-linked, and this hydrogel has permeability, and molecule can exchange through gel partition.Its hollow parts is the nutrient solution containing target cell, and utilize the adherent property of cell, the cell attachment of Dual culture is on the inwall of hollow gelled fibre.
By reference to the accompanying drawings, described the preparation method of the above-mentioned cell co-culture model based on Gel chips by specific embodiment, but the present invention is not only confined to following examples.The present embodiment is the co-culture model of manufacture two kinds of cells, and two kinds of cells adopt scleroblast and osteoclast, observe the change of two kinds of characteristics of cell biology in this co-culture model, inquire into the interaction between scleroblast and osteoclast.Detailed process is as follows:
(1) preparation quality concentration is the sodium alginate soln of 2%: in proportion by biochemical level sodium alginate powder and deionized water mixing, stir, mix, and carry out sterilising treatment with magnetic stirrer;
(2) prepare the calcium chloride solution that concentration is 2%: mixed with deionized water by calcium chloride powder in proportion, stir with magnetic stirrer, mix, and carry out sterilising treatment;
(3) sodium alginate soln of configure in step (1) 2% is poured in the syringe be connected with island shower nozzle exit orifice 12, and syringe is placed on triple channel syringe pump 10;
(4) calcium chloride solution of configure in step (2) 2% is poured into respectively in the syringe be connected with island shower nozzle endoporus 13, endoporus 14, and syringe is placed on triple channel syringe pump 10;
The structure of island shower nozzle, as Fig. 7 and Fig. 8, comprises two endoporus, is endoporus 13, endoporus 14 respectively; Be enclosed in the exit orifice 12 of endoporus 13 and endoporus 14 periphery; The interior passageway 15 be communicated with endoporus 13, endoporus 14; 12 logical exterior passage ways 16 are connected with exit orifice; Interior passageway 15 is connected with respective syringe respectively with exterior passage way 16, fill the calcium chloride solution of configure in step (2) 2% in the syringe be wherein communicated with interior passageway, in the syringe be communicated with exterior passage way, fill the sodium alginate soln of configure in step (1) 2%.
(5) preparation quality concentration is the sodium alginate soln of 4%: in proportion by biochemical level sodium alginate powder and deionized water mixing, stir, mix, and carry out sterilising treatment with magnetic stirrer;
(6) prepare the calcium chloride solution that concentration is 4%: mixed with deionized water by calcium chloride powder in proportion, stir with magnetic stirrer, mix, and carry out sterilising treatment;
(7) in order to form the gel-in-matrix 2 of coated tubular fibre structure, the sodium alginate soln of configure in step (5) 4% is poured in culture dish, utilize Horizontal vibration device sodium alginate is evenly distributed in culture dish and has smooth surface, forming the first layer height is the sodium alginate soln of 4mm.
(8) as shown in Figure 6, the fiber utilizing the type printer based on island shower nozzle 9 to manufacture tubular fibre structure is cultivated body 1 and fiber is cultivated body 1 and is coated in gel: first utilize triple channel syringe pump 10 to control the velocity of flow of the liquid of the exit orifice 12 of island shower nozzle 9, endoporus 13 and endoporus 14, all be set to 1ml/min, obtain tubular fibre structure, next utilizes motion platform 11, the movement velocity regulating platform is 1200mm/min, makes the fiber of formation cultivation body 1 be deposited on the surface of sodium alginate in step (7) with S shape track;
(9) 4% sodium alginate soln configured in step (5) is continued to pour in culture dish, the fiber covering formation is cultivated on body structure, forming second layer height is the sodium alginate soln of 4mm, and with Horizontal vibration device, second layer sodium alginate is evenly distributed in culture dish and has smooth surface;
(10) 4% calcium chloride solution configured in step (6) is ejected in the culture dish in step (9), makes the sodium alginate cross-linking on surface become gel;
(11) total is immersed in the calcium chloride solution of step (6) configures 4%, makes total be cross-linked into the gel structure with some strength completely, finally obtain the Gel chips containing tubular fibre structure.
(12) the triple channel syringe pump in step (3) (only need two passages) is utilized to be expelled to by the nutrient solution containing scleroblast 3 and osteoclast 4 in the adjacent passage of two of Gel chips respectively, injection speed is 4ml/min, leave standstill 12 hours later cell completely adherent, finally obtain the co-culture model of scleroblast based on Gel chips and osteoclast, as shown in Figure 3.
(13) as shown in Figure 4, with dual-channel peristaltic pump 7 continuous circulation in two passages of Gel chips, pour into the nutrient solution of scleroblast and osteoclast, obtain the dynamic Dual culture of scleroblast and osteoclast, wherein be contained in container for storing liquid 5 containing osteoblasts cultivation liquid, hold in container for storing liquid 6 containing Osteoclast culture liquid.
Use the scleroblast based on Gel chips of the present invention and osteoclast co-culture model, observe under inverted microscope, obviously can observe two kinds of cells difference morphologically, and two kinds of cell all energy proliferates under this culture condition, this is that the interaction studied between scleroblast and osteoclast provides the foundation.
Embodiment 2
As shown in Figure 5, also method of the present invention can be adopted to prepare the co-culture model with three passages, one of them channel load has cell A17, channel load has cell B18, a channel load has cell C8, the model of three kinds of co-culture of cells can be formed, when processing the model of three kinds of co-culture of cells, adopt island nozzle structure and Fig. 7 shown in nozzle structure similar, difference is that the quantity of endoporus and interior passageway is 3, and all the other structures are with embodiment 1.
Claims (10)
1. a cell co-culture model, comprise matrix (2), it is characterized in that, be built with fiber cultivation body (1) that matrix (2) outer wall is stretched out at two ends in described matrix (2), this fiber is cultivated body (1) and is made up of gel material; Described fiber is cultivated in body (1) and is provided with two or more passages for co-culture of cells (1a), separate between each passage, each passage (1a) has the entrance (1b) and outlet (1c) that lay respectively at fiber cultivation body (1) two ends; Described gel material is the cross-linking products of sodium alginate and calcium chloride.
2. cell co-culture model according to claim 1, is characterized in that, described matrix (2) is formed by gel material or is made up of polydimethylsiloxane.
3. cell co-culture model according to claim 1, is characterized in that, described fiber is cultivated body (1) and arranged in roundabout shape.
4. a preparation method for cell co-culture model, is characterized in that, comprising:
(1) sodium alginate soln I and calcium chloride solution I is configured;
(2) sodium alginate soln I and calcium chloride solution I is passed in the syringe corresponding from the different passages of island printing head respectively;
(3) body material is configured;
(4) body material that step (3) configures is poured in the container of motion platform, ensure that body material is evenly distributed in a reservoir and has smooth surface simultaneously, form the first layer base material layer;
(5) utilize the island printing head in step (2) to print fiber according to desired trajectory on the first layer base material layer and cultivate body entity, wherein fiber is cultivated body entity periphery and is printed sodium alginate soln I, and fiber cultivates body entity house print for the formation of two or more calcium chloride solution I for the passage of co-culture of cells;
(6) body material that step (3) configures is continued to pour in container, cover the fiber formed and cultivate on body entity;
(7) process is cured to the chip with fiber cultivation body structure that step (6) obtains, obtains many cells co-culture model.
5. the preparation method of cell co-culture model according to claim 4, is characterized in that, the mass percent concentration of described sodium alginate soln I is 1%-3%; The mass percent concentration of described calcium chloride solution I is 2%-5%.
6. the preparation method of cell co-culture model according to claim 4, it is characterized in that, described body material is gel, polydimethylsiloxane, and when body material is gel, described gel is the cross-linking products of sodium alginate soln II and calcium chloride solution II.
7. the preparation method of cell co-culture model according to claim 6, is characterized in that, the mass percent concentration of described sodium alginate soln II is 3%-5%; The mass percent concentration of described calcium chloride solution II is 5%-8%.
8. the preparation method of cell co-culture model according to claim 4, is characterized in that, described island printing head comprises:
Two or more endoporus, quantity and the structure of the quantity of endoporus and structure and passage match;
Be enclosed in the exit orifice of endoporus periphery;
The interior passageway be communicated with respectively with endoporus;
The exterior passage way be communicated with exit orifice;
Described interior passageway and exterior passage way are connected with respective syringe respectively, fill calcium chloride solution I in the syringe be wherein communicated with interior passageway, fill sodium alginate soln I in the syringe be communicated with exterior passage way.
9. the preparation method of cell co-culture model according to claim 8, is characterized in that, the flow velocity of described exit orifice place sodium alginate soln I is 0.5-2ml/min; The flow velocity of described endoporus place calcium chloride solution I is 1-5ml/min.
10. the preparation method of cell co-culture model according to claim 4, is characterized in that, the translational speed of described motion platform is 1000-1600mm/min.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104939946A (en) * | 2015-06-29 | 2015-09-30 | 上海大学 | Method for preparing hollow hydrogel fibers and constructing branch blood vessel unit |
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