CN103966095B - A kind of bio-reactor being applicable to three-dimensional tissue's Cell infusion and cultivating - Google Patents
A kind of bio-reactor being applicable to three-dimensional tissue's Cell infusion and cultivating Download PDFInfo
- Publication number
- CN103966095B CN103966095B CN201410218150.XA CN201410218150A CN103966095B CN 103966095 B CN103966095 B CN 103966095B CN 201410218150 A CN201410218150 A CN 201410218150A CN 103966095 B CN103966095 B CN 103966095B
- Authority
- CN
- China
- Prior art keywords
- box body
- groove
- bio
- applicable
- static pressure
- Prior art date
- 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.)
- Expired - Fee Related
Links
Classifications
-
- 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
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/08—Bioreactors or fermenters specially adapted for specific uses for producing artificial tissue or for ex-vivo cultivation of tissue
-
- 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
- C12M23/38—Caps; Covers; Plugs; Pouring means
-
- 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
- C12M23/42—Integrated assemblies, e.g. cassettes or cartridges
-
- 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
- C12M29/10—Perfusion
-
- 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
- C12M29/18—External loop; Means for reintroduction of fermented biomass or liquid percolate
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Sustainable Development (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Clinical Laboratory Science (AREA)
- Molecular Biology (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The present invention relates to a kind of bio-reactor being applicable to three-dimensional tissue's Cell infusion and cultivating, successively reaction chamber, container for storing liquid are connected with peristaltic pump by tubing; Reaction chamber comprises box body and the lid of the formations such as sidewall, diapire, cut-off equipment, overflow plate, screen bed base, static pressure groove, dredging flow groove, drainage trough, cut-off equipment is gate or weir flow dam and offers some evenly distributed permeable holes along in its longitudinal cross-section same level, static pressure groove sidewall is provided with the overflow port matched with drainage trough, and screen bed base is fixed on the two side of box body.The invention still further relates to a kind of bio-reactor of stepped construction.The invention solves during existing filling type is cultivated the flow field heterogeneity, the uneven problem easily causing necrocytosis of nutrition transmission that exist, be applicable to histiocytic dimensional culture, can improve the quality of tissue engineering product, this reactor has favorable expandability, highly versatile, feature applied widely.
Description
Technical field
The invention belongs to cell culture apparatus field, relate to a kind of bio-reactor being applicable to three-dimensional tissue's Cell infusion and cultivating, the especially bio-reactor of Various Tissues organ Dual culture.
Background technology
The engineering tissue (or claiming three-dimensional constrcts) built under normal circumstances needs to carry out in vitro the cultivation of for some time, could transplant after making its maturation or for research, and training mode the most frequently used is at present the static cultivation method adopting culture plate regularly to change liquid, namely first remove old nutrient solution and add new nutrient solution again, then put into incubator and give quiescent culture, its advantage is the cultivation of applicable short run, changes the liquid cycle and can take corresponding strategy with the difference of tissue, cell.But its shortcoming is apparent: one is the consumption of nutritive medium is that Progressive symmetric erythrokeratodermia declines, and meta-bolites constantly increases, and some physiologicals can be caused to damage to cell; Two is rely on diffusion fashion to provide nutrition, there is the remarkable shortcoming that nutrition supply is limited, even causes the cell of engineering tissue inside to can not get nutrition and dead; Three is periodically change liquid to make nutritive ingredient be cyclic fluctuation, is unfavorable for the homoepitaxial of cell.
In vivo, abundant capillary network framework plays an effective mass transfer channel, for cell provides sufficient oxygen and nutrition.Therefore during vitro culture in order to ensure nutraceutical supply, the method of nutrient solution Efficient Cycle must be set up, there is following strategy at present: roller bottle cultivation, rotating and culturing and filling type are cultivated, wherein the advantage of filling type cultivation is particularly remarkable, on the one hand, because nutrient solution flows through three-dimensional constrcts inside or surrounding continuously, nutritive substance can be overcome and transmit limited shortcoming, even if construction thickness also can obtain reliable nutrition supply more than 500 μm; On the other hand, liquid stream during perfusion culture can produce certain shearing force, provides suitable mechanical stimulus, is conducive to growth and the maturation of construction.(the KalyanaramanBM such as Kalyanaraman, DorothyMS, StevenTB.MediumFlowRateRegulatesViabilityandBarrierFunct ionofEngineeredSkinSubstitutesinPerfusionCulture.TisEng: PartA.2008, 14 (5): 583-93) a kind of biological respinse room has been devised, enter, water outlet pipe accesses culturing room respectively from the both sides, bottom surface of culturing room, the height of culturing room's liquid level is controlled by the height arranging drain pipe, namely excessive inlet pipe flows out higher than liquid after rising pipe by culturing room's liquid level, by carrying out slaking cultivation to the organization engineering skin built, result shows under rate of flooding is 5ml/min or 15ml/min culture condition, histological structure is better than static cultivation, wherein, shown by cell viability detected result, cell viability under the rate of flooding of 5ml/min is significantly better than the cell viability under static cultivation, the proliferative conditions adopting BrdU incorporation methods to detect cell also shows, the hyperplasia situation when rate of flooding 5ml/min or 15ml/min is significantly better than the hyperplasia situation under static cultivation.Contrast only shows a small amount of hyperplasia when also finding rate of flooding 50ml/min, and experimental study demonstrates the barrier that low flow velocity can increase cell viability and maintain epidermis, and be suitable for transplanting, high flow rate produces the degeneration necrosis that high shearing force then can cause cell.
Although existing filling type culture apparatus achieves the circulation of liquid simply, but do not consider the overall condition in flow field, on the one hand, because nutrient solution flow velocity in working cycle is unstable, change is frequent, cannot ensure flowing through the uniform flow field being in low flow velocity in time cultivating tissue engineering product, unfavorable to hyperplasia; On the other hand, the nutrient solution being in edge not easily flows, culture apparatus flow field heterogeneity, and nutrition transmission is uneven, easily causes Growth of Cells uneven; Moreover liquid varies in size to the shearing force that cell applies when flow velocity inequality, when flow velocity is fast shearing force excessive and make cell be vulnerable to damage, be unfavorable for hyperplasia again time flow velocity too low (close to static).The present inventor discloses a kind of filling type bioreactor in Third Military Medical University's journal the 35th volume the 7th phase " design of novel skin organizational project filling type biological respinse room and Preliminary Simulation based on quasistatic Plane Fluid Field ", comprises medium bottle, peristaltic pump, aseptic silica gel pipe, dashpot, overfall dike, culturing room and earial drainage ditch.This bio-reactor is that nutrient solution realizes quasistatic Plane Fluid Field by the mode of overflow, nutrient solution enters dashpot, culturing room is entered through high overfall dike overflow, thus in cultivation, histocyte is supported, then nutrient solution is spilled over to earial drainage ditch through low overfall dike, and last nutrient solution enters into lower one deck reactor dashpot.Utilize this bio-reactor, because nutrient solution exists surface tension, flow into the pressure head instability that the flow velocity of dashpot is non-constant and uneven caused, in addition the earial drainage inclined-plane of high overfall dike can not be in definitely smooth, so when nutrient solution flows through the earial drainage inclined-plane of high overfall dike after dashpot savings potential energy, form the flow liquid that one or several strands are uneven, What is more, define compared with some region on earial drainage inclined-plane under low flow velocity cutout so that substratum enter cultivate section flow field time there is uneven flow field, cause the cell in some region to can not get or obtain little nutritive substance and meta-bolites can not be got rid of in time and occur damage or death, and existing screen cloth is fixed on the diapire of reaction chamber mostly, certain support must be needed to be used for fixing screen cloth, will cause in flow field direction thus, uniform flow field can not be formed on rear side of the pillar of stationary screen rack and also can affect to part cultivation region.
As can be seen here, in filling type is cultivated, make to flow through that to wait to cultivate histiocytic liquid flow field even, flow velocity remains at low flow velocity, to raising tissue engineering product quality, shortens the cultivation time, improves that to cultivate efficiency most important.
Summary of the invention
For the problems referred to above, the technical problem to be solved in the present invention is to provide a kind of bio-reactor being applicable to three-dimensional tissue's cell cultures, and the nutrient solution flow velocity that this bio-reactor can realize Plane Fluid Field is suitable, and even flow field is stablized; Present invention also offers a kind of compact construction, favorable expandability, more optimal stacked bio-reactor.
In order to solve the problems of the technologies described above, be applicable to the bio-reactor of three-dimensional tissue's cell cultures, comprise container for storing liquid, peristaltic pump and at least one comprise the reaction chamber of the rectangular parallelepiped box body of open top and lid composition; A cut-off equipment is fixed in box body one end, forms static pressure groove with box body, and the other end fixes an overflow plate, forms dredging flow groove with box body, is fixed with and cut-off equipment and all discontiguous screen bed base of overflow plate between static pressure groove and dredging flow groove; Screen bed base is fixed on box body, and screen cloth is fixed in the middle of screen bed base; Forming reactions region between cut-off equipment and overflow plate; Reaction chamber is arranged one and be communicated with the inlet opening that also liquid directly can be injected static pressure groove with static pressure groove, the wall of box body arranges one and is communicated with the fluid hole also liquid directly being discharged dredging flow groove with dredging flow groove; Cut-off equipment longitudinally cross section same level offers the some evenly distributed permeable holes liquid in static pressure groove at the uniform velocity evenly being introduced conversion zone; The sidewall of static pressure groove is provided with overflow port, the sidewall of box body is provided with the surplus liquid that static pressure groove can be overflowed through overflow port enters dredging flow groove drainage trough by inflow port; Cut-off equipment is equal to or less than the degree of depth of box body, screen bed base is lower than the height of cut-off equipment, overflow plate is greater than or equal to the height of screen bed base upper screen cloth, the lower edge of overflow port lower than the upper surface of cut-off equipment height and higher than the height of overflow plate upper surface, inflow port and permeable hole upper limb, to form stair-stepping poor position liquid level in static pressure groove, conversion zone and dredging flow groove; Container for storing liquid, peristaltic pump are connected in turn between fluid hole and inlet opening by tubing.
In the present invention, screen bed base is fixed on the two side of box body, cut-off equipment, overflow plate and screen bed base respectively with box body by being connected together, on the face that cut-off equipment, overflow plate contact with box body and the face that contacts with box side wall of screen bed base is provided with step-like clamp, the sidewall of box body with diapire is provided with the engaging groove fastened with clamp.Facilitate dismounting and the cleaning of reaction chamber by being connected together, and compared with sidewall screen bed base being fixed on box body was fixed on diapire with former screen bed base, decreases the stop of screen bed base support to nutrient solution in conversion zone, define uniform flow field.
Overflow port in preferred the present invention comprises U-type groove and rotating by locating shaft of the other setting of U-type groove covers U-type groove and the catch carrying out liquid level in regulating static groove by adjustment U-type groove lower edge height in static pressure groove.By arranging overflow port, can easily in regulating static groove the height of liquid level to control the permeable hole that nutrient solution flows evenly through cut-off equipment.
In the present invention, permeable hole is several evenly distributed identical bar hole, circular port, semi-circular hole or profile holes.By arranging evenly distributed permeable hole, can realize forming uniform runner when substratum enters conversion zone, to realize uniform flow field.
In the present invention, the net plane of screen cloth is provided with the specification mould contour with the support of screen bed base, specification mould can be square, circular, trilateral or polymorphic structure.
In the present invention, preferred overflow plate exceeds screen bed base upper screen cloth height is 0 ~ 4mm.By arranging the overflow plate a little more than screen cloth height, the culture on nutrient solution and screen cloth can be made to form good contact.
In improvement project of the present invention, cut-off equipment is the gate with permeable hole, and permeable hole is positioned at the middle and lower part of gate.
In another improvement project of the present invention, cut-off equipment is the weir flow dam with earial drainage inclined-plane, weir flow dam comprises dam body and lower dam body, the side that lower dam body is adjacent with screen bed base has earial drainage inclined-plane, permeable hole is positioned at the upper dam body on this weir flow dam, this earial drainage inclined-plane and box bottom are 15 ° ~ 25 ° angles, and the height on earial drainage inclined-plane is 1/3 ~ 1/2 of weir flow height of dam degree.
By above-mentioned two kinds of cut-off equipments improved, compared with existing cut-off equipment, nutrient solution by producing at the uniform velocity uniform flow field during permeable hole, and is dismantled, it is convenient to clean.
Another kind of improvement project of the present invention there is provided a kind of stacked box body that can produce in batches, and together this stacked box body consists of the mutual reverse imbedding of box body.
In the present invention, the outside of the upper surface of the sidewall of box body is provided with step-like flange, and the surrounding of the diapire of box body is provided with the object gomphosis groove with the flange tabling of wall upper surface.Multiple box body forms the stacked box body of box body by chimeric docking, and cover cap is combined on the top box body of stacked box body, and the box body on lid and top forms reaction chamber, and the diapire of the box body on top and the box body of adjacent lower form a reaction chamber jointly; The fluid hole of stacked box body middle and upper part box body is connected by the inlet opening of tubing with bottom box body, and the fluid hole of the box body bottom stacked box body is connected with the inlet opening of the reaction chamber at top by tubing, container for storing liquid, peristaltic pump; Stacked box body and lid, tubing, container for storing liquid, peristaltic pump form stacked bio-reactor jointly.
The fluid hole of the box body on the top in preferred above-mentioned improvement project in stacked box body is arranged on the diapire of dredging flow groove, and the static pressure groove being adjacent the box body of chimeric bottom is connected, constitute the inlet opening of bottom box body, so that liquid flows directly in the middle of the static pressure groove of bottom box body from the top box body of stacked box body.By being opened in the fluid hole on diapire, simplify the structure of stacked bio-reactor.
The number of the box body in preferred stacked box body is 2 to 20;
Above-mentioned stacked bio-reactor can also be provided with in the outside of stacked box body and to be made up of casing and case lid and with the aseptic reaction box in the duct that tubing can be made to pass through of matching with top box body static pressure groove and bottom box body fluid hole, laminated cartridge body is arranged in casing, and container for storing liquid and peristaltic pump are positioned at outside reaction box.By arranging reaction box, stacked bio-reactor can be made to react under sterile closed environment.
Said external can also be arranged with cyclic water jacket with the bio-reactor of reaction box outside reaction box, and container for storing liquid and peristaltic pump are positioned at outside cyclic water jacket.So that bio-reactor inside is in temperature constant state.
In technique scheme, when injecting nutrient solution in static pressure groove, nutrient solution is put aside in static pressure groove, regulate the height of overflow port, surplus liquid overflows from overflow port, flows into bottom screen bed base lentamente after ensureing to form steady pressure in static pressure groove with the permeable hole on cut-off equipment, and without the stop of screen bed base support in flow field, make in the middle part of reaction box body, to form uniform stable state Plane Fluid Field, carry out uniform nutrition diffusion; Flow into bottom screen bed base because nutrient solution to form after steady pressure leaching naturally in static pressure groove, therefore the nutrient solution bottom inflow screen bed base remains at lower uniform and stable flow velocity, organ or tissue of the three-dimensional tissue engineering product that this flow velocity can be cultivated for the plan on screen bed base or produce provides the shearing force of uniformity, is conducive to hyperplasia and tissue development.
The present invention is a kind of bio-reactor being applicable to three-dimensional tissue's organ, cell or tissue engineering product perfusion culture, is particularly useful for organization engineering skin or other membranaceous tissues such as cornea, heart valve and Various Tissues organ, cell are carried out to perfusion culture and comprise Dual culture.Its principle of work is: on the material solution filling sieve rack of organ or tissue of the three-dimensional tissue engineering product first cultivated intending or produce, screen bed base is put into the specification that specified shape made by specification mould, takes out specification mould, cover lid, then in static pressure groove, nutrient solution is injected, nutrient solution is put aside in static pressure groove, surplus liquid overflows from overflow port, permeable hole after ensureing to form steady pressure in static pressure groove on cut-off equipment flows to screen bed base, make in the middle part of reaction box body, to form uniform stable state Plane Fluid Field, organ or tissue of the three-dimensional tissue engineering product that plan on screen bed base is cultivated or produced is just by the nutritive medium submergence of flowing, also suitably can reduce liquid level makes three-dimensional tissue to be cultivated on screen bed base be in liquid-gas interface cultivation conditions all the time, for organ or tissue of the three-dimensional tissue engineering product intending research or production provides sufficient nutrient and suitable shearing force, when reacting the height of the liquid level retaining in the middle part of box body higher than overflow plate height, nutrient solution flows into dredging flow groove through overflow plate top, and then flow in container for storing liquid from fluid hole, by peristaltic pump again by indent in meditation for liquor pump, realize enclosed Cyclic culture.
Due to above structure, the present invention has the following advantages: 1. by arranging overflow port and with the cut-off equipment liquid in static pressure groove at the uniform velocity evenly being introduced the permeable hole of conversion zone, when nutrient solution flows through permeable hole, the tension force of easy breakthrough self forms uniform runner, make nutrient solution remains lower stable even velocity of flow when flowing through screen bed base, form the Plane Fluid Field of stable and uniform in reaction box precursor reactant region, be conducive to hyperplasia; 2, bio-reactor of the present invention can singlely independently use, also can multiple series connection or stacked use, there is good extendability, the biological characteristics of three-dimensional tissue's organ can be maintained, ensure that the quality of three-dimensional tissue's organ or product, turn improve the efficiency of fundamental research or production; 3. cut-off equipment of the present invention, specification mould, screen bed base and overflow plate can retract replacing according to actual needs, not only be applicable to the cultivation of organization engineering skin, also film material is applicable to as cornea, heart valve, nerve, blood vessel, mesentery, or the cultivation of other organizer official rank, and dissimilar histoorgan can be cultivated simultaneously, also can be used for the seed cell cultivating adherent growth after removing screen bed base, there is highly versatile, feature applied widely.
Accompanying drawing explanation
Fig. 1 is the schematic perspective view of reaction chamber of the present invention;
Fig. 2 is the schematic perspective view of box body body wall of the present invention;
Fig. 3 is the structural representation of bio-reactor of the present invention;
Fig. 4 is the schematic perspective view of gate in the present invention;
Fig. 5 is the schematic perspective view on weir flow dam in the present invention;
Fig. 6 is the schematic perspective view of screen cloth and screen bed base in the present invention;
Fig. 7 is the structural representation of the stacked bio-reactor of the present invention;
Fig. 8 is the structural representation of the embodiment of the present invention three;
Fig. 9 is the schematic diagram in flow field of the present invention;
In figure: 1, box body; 2, lid; 3, cut-off equipment; 4, gate; 5, weir flow dam; 6, permeable hole; 7, static pressure groove; 8, overflow plate; 9, dredging flow groove; 10, upper dam body; 11, lower dam body; 12, earial drainage inclined-plane; 13, overflow port; 14, locating shaft; 15, catch; 16, drainage trough; 17, inflow port; 18 inlet openings; 19, fluid hole; 20, screen bed base; 21, screen cloth; 22, specification mould; 23 clamps; 24, groove is engaged; 25, flange; 26, gomphosis groove; 27, tubing; 28, container for storing liquid; 29, peristaltic pump; 30, stacked box body; 31, casing; 32, case lid; 33, cyclic water jacket; 34, duct.
Embodiment
Below in conjunction with accompanying drawing, the present invention will be further described.
Embodiment one:
See Fig. 1, Fig. 2, Fig. 3, Fig. 4, Fig. 6, Fig. 9, the present invention is a kind of is applicable to the bio-reactor that histocyte dimensional perfusion cultivates, comprise container for storing liquid 28, peristaltic pump 29 and at least one comprise the reaction chamber that the rectangular parallelepiped box body 1 of open top and lid 2 form; A cut-off equipment 3 is fixed in box body 1 one end, forms static pressure groove 7 with box body 1, and the other end fixes an overflow plate 8, forms dredging flow groove 9 with box body 1, is fixed with and cut-off equipment 3 and all discontiguous screen bed base 20 of overflow plate 8 between static pressure groove 7 and dredging flow groove 9; Screen bed base 20 blocks and on the two side of box body 1, screen cloth 21 is fixed in the middle of screen bed base 20; Cut-off equipment 3 longitudinally cross section same level offers some evenly distributed permeable holes 6 liquid in static pressure groove 7 at the uniform velocity evenly being introduced conversion zone, and permeable hole 6 is several evenly distributed bar hole, circular port, semi-circular hole or profile holes; The sidewall that static pressure groove 7 and box body 1 share is provided with overflow port 13, overflow port 13 comprises U-type groove and rotating by locating shaft 14 of arranging in static pressure groove 7 covers U-type groove and can regulate the catch 15 of U-type groove height and size, and the sidewall of box body 1 is provided with the surplus liquid that static pressure groove 7 can be overflowed through overflow port 13 enters dredging flow groove 9 drainage trough 16 by inflow port 17; Reaction chamber is arranged one and be communicated with the inlet opening 18 that also liquid directly can be injected static pressure groove 7 with static pressure groove 7, the wall of box body 1 arranges one and is communicated with the fluid hole 19 also liquid directly being discharged dredging flow groove 9 with dredging flow groove 9; Cut-off equipment 3 is equal to or less than the degree of depth of box body 1, screen bed base 20 is lower than the height of cut-off equipment 3, overflow plate 8 greater than or equal to the height of screen bed base 20 upper screen cloth 21, the lower edge of overflow port 13 lower than the upper surface of cut-off equipment 3 height and higher than the height of overflow plate 8 upper surface, inflow port 17 and permeable hole 6 upper limb; Container for storing liquid 28, peristaltic pump 29 are connected in turn between fluid hole 19 and inlet opening 18 by tubing 27.
As Fig. 2, shown in Fig. 3, cut-off equipment 3 in the present embodiment is gate 4, peristaltic pump 29 by the cultivation liquid pump in container for storing liquid 28 in the process in static pressure groove 7, inject the nutrient solution higher than permeable hole 6 height, surplus liquid overflows from overflow port 13, pass through overflow groove, inflow port 17 flows in dredging flow groove 9, nutrient solution is by the permeable hole 6 on gate 4, and flow into lentamente bottom screen bed base 20, make in the middle part of box body 1, to form uniform Plane Fluid Field (see Fig. 9), carry out uniform nutrition diffusion, and make the plan organization of production engineering product on screen cloth 21 be subject to the shearing force of uniformity, avoid the necrocytosis caused because nutrient solution nutrition diffusion is uneven or shearing force is too high.
In order to make, the scope of application of the present invention is wider, dismounting cleaning is more convenient, box body 1 sidewall and diapire has engaging groove 24, gate 4, screen bed base 20 and overflow plate 8 have the clamp 23 that can insert and be fixed in described engaging groove 24, gate 4, screen bed base 20 and overflow plate 8 are separable to be fixed on box body 1.Overflow plate 8 is 0 ~ 4mm higher than screen bed base 20 upper screen cloth 21, the liquid level in the middle part of box body 1 can be regulated by replacing overflow plate 8, liquid-gas interface culture environment can not only be provided for organization engineering skin, and can be film material as cornea, heart valve, neural, blood vessel, mesentery etc. provide submerged culture environment, by changing gate 4, screen bed base 20, the catch 15 on overflow plate 8 and slip overflow port 13 side, the shearing force size in adjustable flow field and liquid level, for other three-dimensional tissue's organ as liver, kidney, lung, bone, the tissues such as cartilage and cell are as stem cell, the cultivation of cancer cells, and when removing screen bed base 20, also can be used for the seed cell cultivating adherent growth.
Described screen bed base 20 can be an entirety, also can be multiple separate screen bed base 20, and the spacing of being separated by certain arranges.Contour with the support of screen bed base 20 on the screen cloth 21 that described specification mould 22 is fastened on screen bed base 20, specification mould 22 can be designed as square, circular, trilateral or polymorphic structure according to range of product demand.
(1) by intending research three-dimensional constrcts or intending, on the material solution filling sieve rack 20 of organization of production engineering product, screen cloth 21 being put into the specification that specified shape made by specification mould 22, take out specification mould 22, cover lid 2; Then with the HT speed of 400ml/min to perfusion culture liquid in static pressure groove 7, when the liquid level of nutrient solution is put aside to a certain extent, the catch 15 of slip overflow port 13 is to regulate overflow port 13 height and size, nutrient solution is made to have a small amount of liquid just will overflow from overflow port 13, produce stable static pressure, nutrient solution flows to screen bed base 20 from gate 4 permeable holes 6; Experimental comparison shows, under the low flow velocity condition of this 400ml/min, pours into 2 every day, each half an hour, and cultivate 10 days, hyperplasia is 2.37 times of identical number of days hyperplasia under static culture conditions, can shorten organization engineering skin and cultivate 5 days time.
(2) by intending research three-dimensional constrcts or intending, on the material solution filling sieve rack 20 of organization of production engineering product, screen cloth 21 being put into the specification that specified shape made by specification mould 22, take out specification mould 22, cover lid 2; Then with the medium rate of flooding of 200ml/min to perfusion culture liquid in static pressure groove 7, when the liquid level of nutrient solution is put aside to a certain extent, the catch 15 of slip overflow port 13 is to regulate overflow port 13 height and size, nutrient solution is made to have a small amount of liquid just will overflow from overflow port 13, produce stable static pressure, nutrient solution flows to screen bed base 20 from gate 4 permeable holes 6; Experimental comparison shows, under the low flow velocity condition of this 200ml/min, pours into 2 every day, each half an hour, and cultivate 10 days, hyperplasia is 2.48 times of identical number of days hyperplasia under static culture conditions, can shorten organization engineering skin and cultivate 5 days time.
(3) by intending research three-dimensional constrcts or intending, on the material solution filling sieve rack 20 of organization of production engineering product, screen cloth 21 being put into the specification that specified shape made by specification mould 22, take out specification mould 22, cover lid 2; Then with the Low perfusion speed of 100ml/min to perfusion culture liquid in static pressure groove 7, when the liquid level of nutrient solution is put aside to a certain extent, the catch 15 of slip overflow port 13 is to regulate overflow port 13 height and size, nutrient solution is made to have a small amount of liquid just will overflow from overflow port 13, produce stable static pressure, nutrient solution flows to screen bed base 20 from gate 4 permeable holes 6; Experimental comparison shows, under the low flow velocity condition of this 100ml/min, pours into 2 every day, each half an hour, cultivates 10 days, and hyperplasia is 2.75 times of identical number of days hyperplasia under static culture conditions, shortens tissue engineering product and cultivates 5 days time.
Embodiment two:
As shown in Figure 1, Figure 2, shown in Fig. 3, Fig. 5, Fig. 6, Fig. 9, in above-described embodiment one, the function of gate 4 also can be realized by weir flow dam 5, described weir flow dam 5 comprises dam body 10 and lower dam body 11, the side that lower dam body 11 is adjacent with screen bed base 20 has earial drainage inclined-plane 12, permeable hole is positioned at the upper dam body 10 on this weir flow dam, and this earial drainage inclined-plane 12 is 15 ° ~ 25 ° angles with box bottom, and the height on earial drainage inclined-plane 12 is 1/3 ~ 1/2 of weir flow height of dam degree.Because earial drainage inclined-plane 12 has certain altitude, for avoiding the nutrient solution flowing through earial drainage inclined-plane 12 to cause large impact to screen bed base 20, the flow rate of liquid that the Altitude control on adjustable earial drainage inclined-plane 12 flows to screen bed base 20 is in low flow velocity state.
Embodiment three:
See Fig. 1, Fig. 3, Fig. 7, the outside of the upper surface of the sidewall of the box body 1 of the bio-reactor in first embodiment or second embodiment is provided with step-like flange 25, and the surrounding of the diapire of box body 1 is provided with the object gomphosis groove 26 with flange 25 tabling of wall upper surface.Multiple box body 1 forms stacked box body 30 by reverse imbedding docking, and lid 2 covers on the box body 1 at top; The diapire of box body 1 and the box body 1 of bottom on adjacent stacked box body 30 top form reaction chamber, and box body 1 and the lid 2 at top form reaction chamber; Wherein the fluid hole 19 of box body 1 is positioned on the diapire of box body 1, liquid in the dredging flow groove 9 of top box body 1 also can directly input in the middle of the static pressure groove 7 of bottom box body 1 by the inlet opening 18 that the fluid hole 19 be opened on box body 1 diapire constitutes bottom box body 1, the inlet opening 18 of top box body 1 is positioned on its reaction chamber, and in the middle of the static pressure groove 7 by peristaltic pump 29 liquid that bottom box body 1 fluid hole 19 is discharged directly being pumped into top box body 1; The number of the box body 1 in stacked box body 30 can be 2 to 20, and stacked box body 30 folds bio-reactor with lid 2, tubing 27, container for storing liquid 28, the common constituting layer of peristaltic pump 29.
During use, nutrient solution flows into dredging flow groove 9 after the static pressure groove 7 of top box body 1 flows through cut-off equipment 3, screen bed base 20, overflow plate 8, surplus liquid flows into dredging flow groove 9 from overflow port 13 through drainage trough 16, then the static pressure groove 7 of lower floor's box body 1 is directly injected by fluid hole 19, container for storing liquid 28 is flowed into again from the fluid hole 19 of the box body 1 of bottom, through peristaltic pump 29, the nutrient solution in container for storing liquid 28 is pumped in the static pressure groove 7 of top box body 1 again, realizes the enclosed circulation of nutrient solution.Wherein, the quantity being arranged on the box body 1 at middle part can increase as required or reduce.So circulate, all flow rate of liquid flowing through the middle part of box body 1 are in low flow velocity state all the time, the shearing force that the three-dimensional tissue's organ making the plan organization of production engineering product on screen bed base 20 or intend cultivation can obtain sufficient nutrient and be suitable for, ensure that quality product.
Box body 1 in stacked box body 30 in present embodiment also can use the box body 1 in the middle of as Fig. 1, Fig. 3, and during use, the fluid hole 19 of box body 1 on stacked box body 30 top is also connected by tubing 27 with the inlet opening 18 of adjacent lower box body 1.
Embodiment four:
Accompanying drawing 8 is that the stacked box body 30 in embodiment three is put into aseptic reaction box, reaction box is made up of casing 31 and case lid 32, reaction box offers the duct 34 that tubing 27 can be made to pass through of matching with the inlet opening 18 of stacked box body 30 top box body 1 and the fluid hole 19 of bottom box body 1, stacked box body 30 is arranged in casing 31, and container for storing liquid 28 and peristaltic pump 29 are positioned at outside casing 31.
Can also establish a cyclic water jacket 33 in reaction box periphery, container for storing liquid 28 and peristaltic pump 29 are positioned at outside cyclic water jacket 33.During use, under reaction box can make stacked box body 30 be in gnotobasis, cyclic water jacket can make stacked box body be in temperature constant state, nutrient solution flows at reaction box inner-outer circulation, the flow rate of liquid flowing through the middle part of box body 1 is in low flow velocity state all the time, and the three-dimensional tissue's organ cultivated for the plan organization of production engineering product in reaction box or plan provides sufficient nutrient and suitable shearing force.The cyclic water jacket 33 arranged outside reaction box, has ensured the response behaviour being in constant temperature in reaction box.
The present invention is by arranging gate 4 or weir flow dam 5, match with overflow port 13, no matter under which kind of rate of flooding, nutrient solution can be made in static pressure groove 7 to form stable pressure, comparatively low flow velocity is remained when flowing through screen bed base 20, form uniform stable state Plane Fluid Field in the middle part of box body 1, overcome during existing filling type is cultivated the flow field heterogeneity, the uneven problem easily causing necrocytosis of nutrition transmission that exist, improve quality product, shorten the cultivation time; Also can be used for Dual culture research and the drug screening of three-dimensional constrcts or histoorgan.The present invention also has favorable expandability, highly versatile, feature applied widely, achieves and cultivates the stdn of tissue engineering product, significantly reduce labour intensity, improve production efficiency, has good promotion and application and is worth.
Obviously, all embodiments of foregoing description are a part of embodiments of the present invention, instead of whole embodiments.Based on embodiments of the invention, those of ordinary skill in the art are not making other embodiments all obtained under creative work prerequisite, all belong to the category of the present invention's protection.
Claims (14)
1. be applicable to the bio-reactor that three-dimensional tissue Cell infusion is cultivated, comprise container for storing liquid (28), reaction chamber that the rectangular parallelepiped box body (1) of peristaltic pump (29) and lid (2) and at least one open top forms; Described box body (1) one end is provided with cut-off equipment (3), static pressure groove (7) is formed with box body (1), the other end is provided with overflow plate (8), form dredging flow groove (9) with box body (1), be provided with between static pressure groove (7) and dredging flow groove (9) and cut-off equipment (3) and all discontiguous screen bed base (20 of overflow plate (8)
); Described reaction chamber is provided with the inlet opening (18) being communicated with static pressure groove (7) and also liquid directly can being injected static pressure groove (7), the wall of box body (1) is provided with the fluid hole (19) being communicated with dredging flow groove (9) and also liquid directly can being discharged dredging flow groove (9); The height of described cut-off equipment (3) is equal to or less than the degree of depth of box body (1), and the height of screen bed base (20) is lower than the height of cut-off equipment (3), and the height of overflow plate (8) is greater than or equal to the height of screen bed base (20) upper screen cloth (21); The fluid hole (19) of described reaction chamber, inlet opening (18), container for storing liquid (28) are connected by tubing (27) successively with peristaltic pump (29); It is characterized in that: the sidewall of described static pressure groove (7) is provided with can regulating static groove (7) interior liquid level to form the overflow port (13) of stable static pressure, the sidewall of box body (1) is provided with the surplus liquid that static pressure groove (7) can be overflowed through overflow port (13) enters dredging flow groove (9) drainage trough (16) by inflow port (17); Described cut-off equipment (3) longitudinally cross section same level offers some evenly distributed permeable holes (6) liquid in static pressure groove (7) at the uniform velocity evenly being introduced conversion zone; Regulate the middle flow velocity of liquid of inlet opening (18) and the height of overflow port (13) and size to form stair-stepping poor position liquid level in static pressure groove (7), conversion zone and dredging flow groove (9); Described screen bed base (20) is fixed on the two side of box body (1); The lower edge of described overflow port (13) lower than the upper surface of cut-off equipment (3) height and higher than the height of overflow plate (8) upper surface, inflow port (17) and permeable hole (6) upper limb.
2. a kind of bio-reactor being applicable to three-dimensional tissue's Cell infusion and cultivating according to claim 1, it is characterized in that: on the face that described cut-off equipment (3), overflow plate (8) contact with box body (1) and the face of screen bed base (20) and box body (1) sidewall contact is provided with clamp (23), the sidewall of box body (1) with diapire is provided with the engaging groove (24) fastened with clamp (23).
3. a kind of bio-reactor being applicable to three-dimensional tissue's Cell infusion and cultivating according to claim 1, is characterized in that: described overflow port (13) comprises U-type groove and rotating by locating shaft (14) of the other setting of U-type groove covers U-type groove and the catch (15) carrying out liquid level in regulating static groove (7) by adjustment U-type groove lower edge height in static pressure groove.
4. a kind of bio-reactor being applicable to three-dimensional tissue's Cell infusion and cultivating according to claim 1, is characterized in that: described permeable hole (6) is identical bar hole, circular port, semi-circular hole or other profile holes.
5. a kind of bio-reactor being applicable to three-dimensional tissue Cell infusion and cultivating according to claim 1, is characterized in that: the height of described overflow plate (8) is 0 ~ 4mm higher than the height of the screen cloth (21) on screen bed base (20).
6. a kind of bio-reactor being applicable to three-dimensional tissue's Cell infusion and cultivating according to claim 1, is characterized in that: the gate (4) that described cut-off equipment (3) is rectangular parallelepiped, permeable hole (6) is positioned at the middle and lower part of gate (4).
7. a kind of bio-reactor being applicable to three-dimensional tissue's Cell infusion and cultivating according to claim 1, it is characterized in that: described cut-off equipment (3) is the weir flow dam (5) comprising dam body (10) and lower dam body (11), weir flow dam (5) is rectangular parallelepiped in static pressure groove (7) side, and lower dam body (11) and screen bed base (20) adjacent side are the earial drainage inclined-plane (12) flushed with weir flow dam (5) bottom surface; Described earial drainage inclined-plane (12) and box body (1) bottom surface are 15 ° ~ 25 ° angles, and the height of earial drainage inclined-plane (12) is 1/3 ~ 1/2 of weir flow dam (5) height; Described permeable hole (6) is positioned on dam body (10).
8. a kind of bio-reactor being applicable to three-dimensional tissue's Cell infusion and cultivating according to claim 1, it is characterized in that: the specification mould (22) described screen cloth (21) being provided with cell attachment, specification mould (22) is square, circular, trilateral or polymorphic structure.
9. a kind of bio-reactor being applicable to three-dimensional tissue's Cell infusion and cultivating according to any one of claim 1 to 8, it is characterized in that: multiple box body (1) forms stacked box body (30) by reverse stack docking is fixing, the box body (1) that lid (2) covers at top is gone up and forms top reaction chamber with top box body (1), the fluid hole (19) of the adjacent pack (1) of stacked box body (30) is connected with inlet opening (18), between the fluid hole (19) that container for storing liquid (28) and peristaltic pump (29) are connected to stacked box body (30) bottom box body (1) by tubing (27) and the inlet opening (18) of top reaction chamber.
10. a kind of bio-reactor being applicable to three-dimensional tissue's Cell infusion and cultivating according to claim 9, is characterized in that: in described stacked box body, the quantity of box body is 2 to 20.
11. a kind of bio-reactors being applicable to three-dimensional tissue's Cell infusion and cultivating according to claim 9, it is characterized in that: the outside of the wall upper surface of described box body (1) is provided with step-like flange (25), box body (1) diapire offers can with the gomphosis groove (26) of flange (25) tabling, multiple box body (1) is oppositely and forms stacked box body (30) by flange (25) with docking of gomphosis groove (26).
12. a kind of bio-reactors being applicable to three-dimensional tissue's Cell infusion and cultivating according to claim 9, it is characterized in that: the fluid hole (19) of bottom box body (1) is positioned on the sidewall of box body (1) in described stacked box body (30), the liquid of dredging flow groove (9) can directly be inputted bottom box body (1) static pressure groove (7) by the diapire that the fluid hole (19) of other box bodys (1) is arranged in box body (1).
13. a kind of bio-reactors being applicable to three-dimensional tissue's Cell infusion and cultivating according to claim 9, it is characterized in that: the outside of described stacked box body (30) is provided with the reaction box be made up of casing (31) and case lid (32), reaction box offers the duct (34) that tubing (27) can be made to pass through, container for storing liquid (28) and peristaltic pump (29) are positioned at outside reaction box.
14. a kind of bio-reactors being applicable to three-dimensional tissue's Cell infusion and cultivating according to claim 13, it is characterized in that: be arranged with cyclic water jacket (33) outside described reaction box, container for storing liquid (28) and peristaltic pump (29) are positioned at cyclic water jacket (33) outward.
Priority Applications (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410218150.XA CN103966095B (en) | 2014-05-22 | 2014-05-22 | A kind of bio-reactor being applicable to three-dimensional tissue's Cell infusion and cultivating |
| US15/328,460 US9902929B2 (en) | 2014-05-22 | 2015-05-20 | Bioreactor for three-dimensional tissue perfusion culture |
| PCT/CN2015/079330 WO2015176653A1 (en) | 2014-05-22 | 2015-05-20 | Bioreactor for three-dimensional tissue perfusion culture |
| EP15795352.2A EP3147349B1 (en) | 2014-05-22 | 2015-05-20 | Bioreactor for three-dimensional tissue perfusion culture |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410218150.XA CN103966095B (en) | 2014-05-22 | 2014-05-22 | A kind of bio-reactor being applicable to three-dimensional tissue's Cell infusion and cultivating |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103966095A CN103966095A (en) | 2014-08-06 |
| CN103966095B true CN103966095B (en) | 2016-01-20 |
Family
ID=51236104
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410218150.XA Expired - Fee Related CN103966095B (en) | 2014-05-22 | 2014-05-22 | A kind of bio-reactor being applicable to three-dimensional tissue's Cell infusion and cultivating |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103966095B (en) |
Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015176653A1 (en) * | 2014-05-22 | 2015-11-26 | 第三军医大学第三附属医院 | Bioreactor for three-dimensional tissue perfusion culture |
| US11186810B2 (en) | 2016-03-16 | 2021-11-30 | Shimadzu Corporation | Cell culture vessel |
| CN105754856B (en) * | 2016-04-12 | 2018-06-22 | 中国航天员科研训练中心 | A kind of drawer type three-dimensional class blood vessel tissue culture organ chip and its construction method |
| CN105950468B (en) * | 2016-05-23 | 2018-03-02 | 西安交通大学 | A kind of bioreactor for perfusion and the bioreactor for infiltration |
| CN115044471A (en) | 2016-08-27 | 2022-09-13 | 三维生物科技有限公司 | Bioreactor |
| CN106940333A (en) * | 2017-04-17 | 2017-07-11 | 山西医科大学 | The outer Electrophysiology research perfusion system of isolated cells |
| CN108060081A (en) * | 2018-02-05 | 2018-05-22 | 中山大学 | A kind of multi-layer planar filling type bioreactor |
| CN108749274B (en) * | 2018-04-30 | 2022-10-04 | 浙江聚众柔印科技有限公司 | Traditional Chinese medicine water dispersing device of flexible plate making machine |
| KR102381523B1 (en) * | 2020-10-15 | 2022-03-31 | 주종일 | Three-dimensional cell culture chip with automatic supplying cell culture media |
| CN113512497B (en) * | 2021-08-12 | 2024-05-28 | 浙江大学 | 3D printing culture tank, culture system and detection system for in-vitro intestinal living body culture and application |
| CN114574333B (en) * | 2022-02-23 | 2023-01-10 | 南京邮电大学 | Nerve cell culture dish |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1560225A (en) * | 2004-03-01 | 2005-01-05 | 华东理工大学 | Pouring type biologic reactor system in vitro constructure of dual active skin |
| CN203855588U (en) * | 2014-05-22 | 2014-10-01 | 中国人民解放军第三军医大学第三附属医院 | Bioreactor suitable for three-dimensional tissue cell perfusion culture |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2714886B1 (en) * | 2011-05-30 | 2020-01-22 | Atech Partners Ltd. | Bioreactor system for continuous cell cultivation |
-
2014
- 2014-05-22 CN CN201410218150.XA patent/CN103966095B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1560225A (en) * | 2004-03-01 | 2005-01-05 | 华东理工大学 | Pouring type biologic reactor system in vitro constructure of dual active skin |
| CN203855588U (en) * | 2014-05-22 | 2014-10-01 | 中国人民解放军第三军医大学第三附属医院 | Bioreactor suitable for three-dimensional tissue cell perfusion culture |
Non-Patent Citations (2)
| Title |
|---|
| Medium flow rate regulates viability and barrier function of engineered skin substitutes in perfusion culture;Balaji kalyanaraman 等;《Tissue Engineering:Part A》;20081231;第14卷;第583－593页 * |
| 基于准静态平面流场的新型皮肤组织工程灌注式生物反应室设计与初步模拟;王洋 等;《第三军医大学学报》;20130415;第35卷(第7期);第622－626页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103966095A (en) | 2014-08-06 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103966095B (en) | A kind of bio-reactor being applicable to three-dimensional tissue's Cell infusion and cultivating | |
| WO2015176653A1 (en) | Bioreactor for three-dimensional tissue perfusion culture | |
| CN203855588U (en) | Bioreactor suitable for three-dimensional tissue cell perfusion culture | |
| CN106232801A (en) | Automatic cytological is cultivated and harvesting apparatus | |
| CN104611225A (en) | Bioreactor for constructing tissue engineering liver by using in-vitro circulatory perfusion | |
| CN103328625A (en) | Bioreactor | |
| CN206783688U (en) | A kind of multilevel cell amplification system | |
| JP2022501003A (en) | Fluid device with microplate with interconnected wells | |
| US20170145361A1 (en) | Bioreactor with separate co2 supply | |
| CN102575216A (en) | Submerged perfusion bioreactor | |
| CN101824382B (en) | Tissue engineering myocardium bioreactor constructed by pouring, perfusion and pulsation combination | |
| US20160281045A1 (en) | Cell Culture System | |
| CN105695328A (en) | Co-culture device for cell culture and cell-cell interaction and using method of co-culture device | |
| CN204779609U (en) | Extracorporal circulatory system perfusion founds bioreactor of organizational project liver | |
| CN103614296B (en) | Double-chamber three-dimensional pouring bioreactor system | |
| CA2387549A1 (en) | Method for culturing and/or treating cells | |
| US20120315694A1 (en) | Cell culture unit and cell culture device including the same | |
| CN111269830B (en) | Multi-organ chip based on microfluidic technology and application thereof | |
| WO2023246341A1 (en) | Cell culture method, cell culture container, and cell culture device | |
| CN205473829U (en) | Interact's culture apparatus between cell culture and cell | |
| CN107287117B (en) | Device for large-scale culture and freezing of hepatocytes | |
| IL267759B2 (en) | Perfusion device | |
| CN102010823A (en) | Cell culture solution flow supply device | |
| CN204162724U (en) | A kind of perfusion cap cultivated for biological sample filling type | |
| CN102703325A (en) | Method for producing chlamydomonas at low cost |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160120 |