+

WO2012032646A1 - Dispositif de culture cellulaire et procédé de culture cellulaire - Google Patents

Dispositif de culture cellulaire et procédé de culture cellulaire Download PDF

Info

Publication number
WO2012032646A1
WO2012032646A1 PCT/JP2010/065585 JP2010065585W WO2012032646A1 WO 2012032646 A1 WO2012032646 A1 WO 2012032646A1 JP 2010065585 W JP2010065585 W JP 2010065585W WO 2012032646 A1 WO2012032646 A1 WO 2012032646A1
Authority
WO
WIPO (PCT)
Prior art keywords
cells
cell culture
intermediate layer
culture
cell
Prior art date
Application number
PCT/JP2010/065585
Other languages
English (en)
Japanese (ja)
Inventor
藤山　陽一
陽一 田川
Original Assignee
株式会社島津製作所
国立大学法人東京工業大学
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by 株式会社島津製作所, 国立大学法人東京工業大学 filed Critical 株式会社島津製作所
Priority to JP2012532811A priority Critical patent/JP5700460B2/ja
Priority to PCT/JP2010/065585 priority patent/WO2012032646A1/fr
Publication of WO2012032646A1 publication Critical patent/WO2012032646A1/fr

Links

Images

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
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/16—Microfluidic devices; Capillary tubes
    • 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
    • C12M25/02—Membranes; Filters

Definitions

  • the present invention relates to a cell culture device and a cell culture method using the same.
  • Cell culture is generally performed in a state where cells and a liquid medium are contained in a container such as a petri dish.
  • a container such as a petri dish.
  • the application of microdevices manufactured by microfabrication technology has been promoted in the medical and biotechnology research fields, and cell culture using such microdevices has been promoted. (For example, refer to Patent Document 1).
  • a microdevice for cell culture (cell culture device) is formed by forming a culture chamber and a microchannel inside a flat substrate, and performs cell culture by accommodating cells and a medium in the culture chamber. The medium is exchanged using the microchannel.
  • hepatic parenchymal cells constituting 70-80% of the liver and non-parenchymal cells such as sinusoidal endothelial cells are regularly arranged. Signal transduction and fluid circulation between these cells are It plays an important role in maintaining normal liver function.
  • the hepatic parenchymal cells and sinusoidal endothelial cells are arranged side by side, but an extracellular matrix called a dyssel cavity exists between them, and the cell line of hepatic parenchymal cells and the cell line of sinusoidal endothelial cells There is no direct contact. Further, outside the respective cell rows, the hepatic parenchymal cell side is a bile duct and the sinusoidal endothelial cell side is a blood vessel, and drugs and nutrients are supplied from the blood vessel side and discharged to the bile duct side.
  • liver function In order to establish an artificial liver system with liver function, regular arrangement of these cell types and construction of an extracellular matrix are important. However, hepatocytes are difficult to culture for a long period of time, and moreover, it is more difficult to form a tissue close to the liver in vivo. Therefore, an artificial system that can maintain a stable liver function over a long period of time has not yet been constructed.
  • Patent Document 1 In connection with the construction of such an artificial liver system, the present inventors have proposed a biodevice using two types of living cells (Patent Document 1). This uses a porous membrane as an alternative to the extracellular matrix, and by fixing and culturing different cells on both sides of the membrane, signal transmission and substance exchange between the cells on both sides via the porous membrane Or it is made to move.
  • Such a culture device is epoch-making in that it can realize a configuration close to that of a living body as compared with the case of culturing one kind of cell alone, but there is room for further improvement in terms of long-term maintenance of cell functions. was there.
  • the present invention has been made in view of the above points.
  • the object of the present invention is to use a cell culture device capable of maintaining a stable function over a long period of time using two types of biological cells, and the same. It is to provide a cell culture method.
  • the cell culture device which has been made to solve the above problems, a) an intermediate layer consisting of a porous membrane and coated with a scaffold for cell culture; b) a first culture chamber and a second culture chamber formed by partitioning the space inside the device with the intermediate layer; c) a first limiting layer that covers a part of one surface of the intermediate layer and limits a region where the intermediate layer is in contact with the liquid flowing in the first culture chamber; d) a second limiting layer that covers a part of the other surface of the intermediate layer and limits the region where the intermediate layer is in contact with the liquid flowing in the second culture chamber; It is characterized by having.
  • the cell culture device has two culture chambers partitioned by an intermediate layer, and can culture different types of cells simultaneously in each culture chamber.
  • the heterogeneous cells are fixed to both sides of the intermediate layer coated with the scaffold, and signal transmission between the cells and exchange of substances can be performed through the porous membrane constituting the intermediate layer.
  • the cells are suspended in a medium and the suspension is injected into the culture chamber by a syringe or the like.
  • the injected cells are biased to adhere to the vicinity of the entrance of the culture chamber, making it difficult to spread the cells throughout the culture chamber.
  • the cell suspension and the intermediate layer are introduced when cells are introduced into the device by providing the first limited layer and the second limited layer.
  • porous film one made of polycarbonate can be suitably used, but is not limited to this, and is made of polytetrafluoroethylene, mixed cellulose, polyethylene terephthalate (PET), glass fiber, polyether, or fluorine resin. And various types such as cellulose-based, nylon-based, and ceramic-based porous membranes can be used.
  • the porous membrane must be such that it does not pass cells and does not hinder the efficiency of substance exchange and signal transmission between cells. Therefore, it is desirable to use a porous membrane having a thickness of 1 mm or less and an average pore diameter of 0.1 ⁇ m to 10 ⁇ m.
  • the first limited layer and the second limited layer have a plurality of openings that expose the intermediate layer to the liquid flowing through the culture chambers, and the opening area per one opening is 0. It is desirable that the thickness be 0.01 mm 2 to 10 mm 2 .
  • the opening area is smaller than this, the number of cells that can be fixed to the intermediate layer with one opening is reduced, and if it is larger than this, it is difficult to obtain the effect of limiting the adhesion region.
  • the thickness of the limiting layer is suitably 1 mm or less. When larger than this, since the said opening part becomes deep, the efficiency of the liquid exchange around a cell in culture
  • the intermediate layer is formed by stacking a plurality of porous membranes.
  • E-cad-Fc a fusion protein of an extracellular region of E-cadherin and an Fc fragment of an antibody IgG molecule
  • E-cad-Fc a fusion protein of an extracellular region of E-cadherin and an Fc fragment of an antibody IgG molecule
  • polymers containing collagen, fibronectin, laminin, PVLA (poly-Np-vinylbenzyl-D-lactone amide) and the like can be used.
  • the cell culture method according to the present invention is a cell culture method using the cell culture device according to the present invention, wherein different types of cells are fixed to one surface and the other surface of the intermediate layer and cultured. It is characterized by doing.
  • the different cells are typically hepatic parenchymal cells and sinusoidal endothelial cells, but the cell device according to the present invention is not limited to these and can be used for culturing various cells.
  • the cell culture device and the cell culture method according to the present invention it is possible to easily realize an environment close to the living body in the cell culture using two types of living cells, and stable cells.
  • the function can be obtained over a long period of time.
  • FIG. 2 is a cross-sectional view taken along arrow XX in FIG. 1.
  • FIG. which shows the evaluation result of the testosterone hydroxylation ability of the cultured cell in Experimental example 2.
  • FIG. 1 is a plan view of the cell culture device according to the present embodiment
  • FIG. 2 is a cross-sectional view taken along the line XX of FIG. 1
  • FIG. 3 is an exploded perspective view of the cell culture device according to the present embodiment.
  • FIG. 1 for the sake of explanation, a part of the internal structure is shown through.
  • the cell culture device 10 has a configuration in which two porous films 14 and 24, limited layers 13 and 23, and seal layers 12 and 22 are sandwiched between two substrates 11 and 21, respectively. It has become.
  • the dimension and material of each part described below are an example to the last, and this invention is not limited to this.
  • the substrates 11 and 21 are made of PDMS (manufactured by Toray Dow Corning, SILPOT184).
  • a first culture chamber 11a composed of a rectangular recess having a length of 16 mm and a width of 2 mm, and groove-shaped channels 11b and 11c extending from the vicinity of both ends of the first culture chamber 11a. Is formed.
  • the first culture chamber 11a and the channels 11b and 11c all have a depth of 0.1 mm and can be formed by molding.
  • the ends of the two flow paths 11b and 11c communicate with a liquid introduction port 11d or a liquid discharge port 11e each formed of a through hole extending in the thickness direction of the substrate 11.
  • a second culture chamber 21a, flow paths 21b and 21c, a liquid inlet 21d, and a liquid outlet 21e are formed in the substrate 21 in the same manner.
  • the limiting layers 13 and 23 are made of a metal mask made of SUS having a thickness of 0.1 mm, and the central portion thereof has a diameter of 0.5 mm in a region corresponding to the first culture chamber 11a or the second culture chamber 21a, respectively. 30 through-holes 13a and 23a are formed.
  • the through holes 13a and 23a correspond to the openings in the present invention.
  • the shape and number of the openings are not limited to the above, and can be various depending on the type of cells to be cultured and the purpose of the culture.
  • the limiting layers 13 and 23 are made of a material that is difficult to adhere to cells to be cultured (at least those having lower cell adhesion than the porous membranes 14 and 24 coated with a scaffold). desirable. As such a material, in addition to the metal such as SUS, various resins can be used.
  • the seal layers 12 and 22 are made of silicon rubber having a thickness of 0.1 mm, and the length and width are almost the same as those of the substrates 11 and 21.
  • a rectangular hole having substantially the same length and width as the limiting layers 13 and 23 is formed in the center of the sealing layers 12 and 22, and the limiting layer 13 or the limiting layer 23 is fitted into the rectangular holes.
  • the porous membranes 14 and 24 are punched into a rectangle having a width of 2 mm and a length of 14 mm.
  • polycarbonate membrane filters (Millipore Corp., Isopore HTTP04700, filter pore diameter 0.4 ⁇ m, thickness 7-22 ⁇ m) were used as the porous membranes 14, 24.
  • superposed these porous membranes 14 and 24 is equivalent to the intermediate
  • the porous membranes 14 and 24 are each coated with a scaffold according to the type of cells to be cultured.
  • the porous membrane 14 on the first culture chamber 11a side was coated with E-cad-Fc
  • the porous membrane 24 on the second culture chamber 21a side was coated with type I collagen.
  • the method of coating the scaffold material is not particularly limited. For example, after the porous films 14 and 24 are dipped and taken out from the liquefied scaffold material, the scaffold material adhered to the porous films 14 and 24 is solidified. Can be considered.
  • Such coating with a scaffold may be performed at the manufacturing stage of the cell culture device, or may be performed by a user who has purchased the cell culture device.
  • each of the above layers is sterilized and the porous membranes 14 and 24 are coated with a scaffold, and then the substrate 21, the seal layer 22, the limiting layer 23, and the porous membrane 24.
  • the porous film 14, the limiting layer 13, the seal layer 12, and the substrate 11 are superposed in this order.
  • the substrates 11 and 21 are configured such that the surface on which the first culture chamber 11a is formed faces the surface on which the second culture chamber 21a is formed.
  • the limiting layers 13 and 23 are fitted into the holes in the center of the sealing layers 12 and 22, the sealing layer 12 and the limiting layer 13, and the sealing layer 22 and the limiting layer 23 respectively. It will be arranged at the same position in the thickness direction.
  • the substrate 11 and the seal layer 12, the substrate 21 and the seal layer 22, and the seal layers 12 and 22 are adsorbed by the self-adsorption property of the silicon rubber constituting the seal layers 12 and 22.
  • the Therefore, the substrate 11 and the substrate 21 can be fixed to each other with the porous films 14 and 24 and the limiting layers 13 and 23 sandwiched therebetween.
  • hepatic parenchymal cells are introduced from the liquid inlet 11d into the first culture chamber 11a, and sinusoidal endothelial cells are introduced from the liquid inlet 21d into the second culture chamber 21a.
  • Each cell is fixed to the surface of the porous membranes 14 and 24, respectively.
  • the cell culture device 10 is first placed so that the first culture chamber 11a side is facing upward, the liver parenchymal cells are suspended in the medium, and the suspension is injected into the first culture chamber 11a with a syringe or the like. To do.
  • the injected cells sink by gravity and adhere to the porous membrane 14.
  • the cell culture device 10 is placed so that the second culture chamber 21a side is up, and sinusoidal endothelial cells are injected into the second culture chamber 21a in the same manner and adhered to the porous membrane 24.
  • Part of the porous membranes 14 and 24 is masked by the limiting layers 13 and 23 so that the liquid in the first culture chamber 11a or the second culture chamber 21a can be contacted only at the through holes 13a and 23a. It has become. For this reason, some of the cells introduced together with the medium from the liquid inlets 11d and 21d as described above enter the through holes 13a and 23a near the upstream of the culture chamber and adhere to the porous membranes 14 and 24. The remaining cells float in the medium or fall on the surfaces of the limiting layers 13 and 23. Since the limiting layers 13 and 23 are less likely to adhere cells than the porous membranes 14 and 24, these cells ride on the flow of the medium and flow downstream of the culture chambers 11a and 21a.
  • one of the through-holes 13a and 23a enters, and adheres to the porous films 14 and 24 there.
  • the cells injected into the device are cultured as in the past. It is possible to prevent the cells from being biased to adhere to the vicinity of the entrance of the chamber and to spread the cells throughout the culture chamber. Further, since the cells gather inside the through holes 13a and 23a, stable cell functions can be exhibited even when culturing cells that form a tight junction with the surrounding cells.
  • FIG. 4 shows an outline of a cell culture system using the cell culture device 10 according to the present example.
  • This is a combination of the cell culture device 10 and a liquid feeding mechanism for continuously feeding a medium to the cell culture device 10.
  • the liquid feeding mechanism is configured to operate the medium storage units 31 and 41, the medium supply pipes 32 and 42, the medium discharge pipes 33 and 43, the waste liquid storage units 34 and 44, the liquid feed pumps 35 and 45, and the liquid feed pumps 35 and 45.
  • the control part 50 which controls is provided.
  • One end of each of the medium supply pipes 32 and 42 is inserted into the medium reservoirs 31 and 41, and the other end is inserted into the liquid introduction ports 11d and 21d of the cell culture device 10, respectively.
  • One ends of the medium discharge pipes 33 and 43 are respectively inserted into the liquid discharge ports 11e and 21e of the cell culture device 10, and the other ends are respectively inserted into the waste liquid storage portions 34 and 44.
  • a medium suitable for culturing hepatocytes is stored in the medium reservoir 31, and a medium suitable for culturing sinusoidal endothelial cells is stored in the medium reservoir 41.
  • the medium stored in the medium storage unit 31 is sucked by the liquid feed pump 35 and sent to the cell culture device 10 through the medium supply pipe 32.
  • the medium supplied to the cell culture device 10 passes through the first culture chamber 11a, and is discharged to the waste liquid storage part 34 through the medium discharge pipe 33 connected to the liquid discharge port 11e.
  • the culture medium stored in the culture medium storage unit 41 is sucked by the liquid feed pump 45, passes through the culture medium supply pipe 42, passes through the second culture chamber 21 a, and is discharged to the waste liquid storage part 44 through the culture medium discharge pipe 43. Is done.
  • the culture medium is exposed to the outside from the liquid inlets 11d and 21d or the liquid outlets 11e and 21e. Although it does not leak out, in order to prevent liquid leakage more reliably, it is desirable to fix the cell culture device 10 by sandwiching it from above and below with an appropriate jig.
  • the porous membranes 14 and 24 can realize an ordered arrangement of cells instead of the extracellular matrix, and signal transmission and substance exchange between cells can be realized. It can be secured. Furthermore, the culture media in the first culture chamber 11a and the second culture chamber 21a play roles similar to bile and blood, respectively, and can exhibit functions close to the living body such as metabolism and drug uptake / discharge.
  • the cell culture device 10 by restricting the region to which the cells can be adhered by the limiting layers 13 and 23, the cells are biased and adhered near the entrances of the culture chambers 11a and 21a.
  • the cells can be adhered to the culture chambers 11a and 21a in a desired pattern suitable for function expression.
  • two porous membranes 14 and 24 it becomes possible to use different scaffolding materials according to the types of cells cultured in the first culture chamber 11a and the second culture chamber 21a. As a result, an environment closer to the living body can be established inside the device, and cell culture can be performed in a state where expression of liver function is maintained for a long period of time.
  • the substrates 11 and 21 are fixed to each other by the seal layers 12 and 22.
  • the seal layers 12 and 22 are not provided and the self-adsorption property of the PDMS constituting the substrates 11 and 21 is not provided.
  • the substrates 11 and 21 may be fixed to each other. In this case, in order to obtain stronger adhesion, it is desirable to activate and bond the bonding surfaces of the substrates 11 and 21 with oxygen plasma or ultraviolet rays.
  • the cell culture device according to the present invention can be used as an artificial organ such as the above-described hybrid type artificial liver, and can also be used as a reaction container when performing a drug metabolism test using cells.
  • an artificial organ such as the above-described hybrid type artificial liver
  • the cell culture device of the present invention can also be used for culturing other cells.
  • Example 1 In the cell culture device 10 as shown in FIGS. 1 to 3, cell culture was performed by changing the material of the porous membranes 14 and 24, and the difference in liver function was evaluated.
  • the porous membranes 14 and 24 three types of porous membranes each made of polycarbonate, polytetrafluoroethylene, or mixed cellulose were used, and each was coated with type I collagen and used in the experiment.
  • the liver function on the first day of culture was evaluated by examining the hydroxylation ability of testosterone by CYP (cytochrome P450). Specifically, first, about 3 ⁇ 10 3 liver parenchymal cells were seeded in the first culture chamber, and about 3 ⁇ 10 3 sinusoidal endothelial cells were seeded in the second culture chamber. Then, a culture medium containing testosterone at a concentration of 0.25 mM is poured into the first culture chamber, a culture medium containing no testosterone is poured into the second culture chamber, and the culture is conducted to the medium (waste liquid) discharged from the first culture chamber. The concentration of testosterone hydroxide contained was measured by HPLC. Each medium was continuously fed to each culture chamber at a flow rate of 40 ⁇ L / hr.
  • the hydroxylation pattern (that is, the abundance ratio of various hydroxylated testosterone in the waste liquid) in the case of using the above three kinds of porous membranes was compared with the hydroxylation pattern by liver microsomes.
  • the hydroxylation pattern by liver microsomes was determined by adding the same medium as that introduced into the first culture chamber to the liver microsome fraction, incubating for 24 hours, collecting the supernatant, and containing various hydroxides by HPLC. Determined by measuring the amount.
  • 16 ⁇ -OHT, 2 ⁇ -OHT, 16 ⁇ -OHT, 6 ⁇ -OHT, and 7 ⁇ -OHT mean testosterone in which the 16 ⁇ , 2 ⁇ , 16 ⁇ , 6 ⁇ , and 7 ⁇ positions are hydroxylated, respectively.
  • a porous membrane made of polycarbonate was used, a hydroxylation pattern closest to liver microsomes was obtained.
  • the cell culture device used in this example uses a polycarbonate porous membrane as the porous membranes 14 and 24, and a scaffold material to be coated on the porous membrane 14 on the liver parenchymal cell side (that is, the first culture chamber 11a side) is used. Except for the changed points, the second embodiment is the same as the first embodiment.
  • As the liver parenchymal cell side coating six types of type I collagen, type IV collagen, fibronectin, laminin, PVLA, and E-cad-Fc were used.
  • hepatocellular parenchymal cells were seeded on the first culture chamber side and about 3 ⁇ 10 3 sinusoidal endothelial cells were seeded on the second culture chamber side, and cell culture was performed. Hydroxylation ability and urea synthesis ability (capability of decomposing ammonia to synthesize urea) on the first day, the fourth day, and the seventh day of the culture were evaluated.
  • the testosterone hydroxylation ability was discharged from the first culture chamber by culturing by flowing a medium containing testosterone in the first culture chamber and a medium not containing testosterone in the second culture chamber, as in Experimental Example 1 above.
  • the concentration of testosterone hydroxide contained in the medium was evaluated by measuring with HPLC.
  • the ability to synthesize urea was obtained by flowing a medium containing NH 4 Cl at a concentration of 2.0 mM in the first culture chamber, and a medium not containing NH 4 Cl flowing in the second culture chamber, On day 7, the urea concentration in the medium discharged from the first culture chamber was evaluated by measuring by HPLC. In each case, each medium was continuously fed to each culture chamber at a flow rate of 40 ⁇ L / hr.
  • FIG. 7 shows the evaluation results of testosterone hydroxylation ability
  • FIG. 8 shows the evaluation results of urea synthesis ability.
  • type I collagen that is, the same as the sinusoidal endothelial cell side. Testosterone hydroxylating ability was higher than when the coating was applied.
  • the porous membrane is made of type IV collagen, fibronectin, laminin, PVLA, or E-cad-Fc rather than the case where the porous membrane on the hepatocyte side is coated with type I collagen.
  • the urea synthesis ability was maintained for a longer period of time (that is, the urea synthesis ability on the seventh day of culture increased).

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Biomedical Technology (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Immunology (AREA)
  • Dispersion Chemistry (AREA)
  • Clinical Laboratory Science (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

L'invention concerne un dispositif de culture cellulaire, comprenant : une couche intermédiaire consistant en deux membranes poreuses (14, 24) ; une première chambre de culture (11a) et une seconde chambre de culture (21a), lesdites chambres de culture étant cloisonnées par la couche intermédiaire ; et des couches limitantes (13, 23) qui sont disposées sur les deux côtés de la couche intermédiaire et dotées de multiples ouvertures (13a, 23a) pour permettre l'exposition de la couche intermédiaire à un liquide s'écoulant entre les chambres de culture (11, 21a). Puisque la couche intermédiaire est constituée des deux membranes poreuses (14, 24), différents échafaudages qui sont adaptés pour le type de cellules à mettre en culture, peuvent être appliquées en revêtement sur les deux côtés de la couche intermédiaire. Puisque les zones auxquelles des cellules peuvent adhérer sont limitées par les couches limitantes (13, 23), les cellules peuvent être fixées à la couche intermédiaire selon un motif souhaité qui est adapté à l'expression de leur fonction. Ainsi, un environnement plus proche de l'environnement in vivo peut être construit à l'intérieur du dispositif et les cellules peuvent être mises en culture sur une longue période de temps tout en conservant leur fonction cellulaire.
PCT/JP2010/065585 2010-09-10 2010-09-10 Dispositif de culture cellulaire et procédé de culture cellulaire WO2012032646A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP2012532811A JP5700460B2 (ja) 2010-09-10 2010-09-10 細胞培養デバイス及び細胞培養方法
PCT/JP2010/065585 WO2012032646A1 (fr) 2010-09-10 2010-09-10 Dispositif de culture cellulaire et procédé de culture cellulaire

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/JP2010/065585 WO2012032646A1 (fr) 2010-09-10 2010-09-10 Dispositif de culture cellulaire et procédé de culture cellulaire

Publications (1)

Publication Number Publication Date
WO2012032646A1 true WO2012032646A1 (fr) 2012-03-15

Family

ID=45810267

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2010/065585 WO2012032646A1 (fr) 2010-09-10 2010-09-10 Dispositif de culture cellulaire et procédé de culture cellulaire

Country Status (2)

Country Link
JP (1) JP5700460B2 (fr)
WO (1) WO2012032646A1 (fr)

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2788469A1 (fr) * 2011-12-07 2014-10-15 A4TEC - Association for The Advancement of Tissue Engineering and Cell Based Technologies and Therapies Procédé à une seule étape et dispositif de génération de substituts de tissu tubulaire stratifié
WO2015159333A1 (fr) * 2014-04-14 2015-10-22 株式会社島津製作所 Dispositif de culture cellulaire, système de culture cellulaire et procédé de culture cellulaire
WO2015169287A1 (fr) * 2014-05-08 2015-11-12 Universitätsklinikum Jena Procédé et dispositifs pour la production in vitro d'ensembles de couches cellulaires
JP2016214148A (ja) * 2015-05-20 2016-12-22 住友電気工業株式会社 細胞培養担体及びこれを備える細胞シート
WO2017096192A1 (fr) 2015-12-04 2017-06-08 President And Fellows Of Harvard College Dispositifs de simulation d'une fonction d'un tissu hépatique et leurs procédés d'utilisation et de fabrication
JP6205507B1 (ja) * 2016-05-19 2017-09-27 光次 斉藤 培養装置および培養方法、並びにこの培養方法により製造された培養臓器
JP2019518443A (ja) * 2016-05-11 2019-07-04 ユニヴェルシテ グルノーブル アルプ 細胞共培養チップ及びその製造に関するプロセス
WO2019168118A1 (fr) * 2018-03-02 2019-09-06 富士フイルム株式会社 Dispositif à canaux microfluidiques
US10472612B2 (en) 2011-02-28 2019-11-12 President And Fellows Of Harvard College Cell culture system
JP2020010641A (ja) * 2018-07-18 2020-01-23 株式会社島津製作所 細胞培養装置および細胞播種方法
CN111205959A (zh) * 2018-11-21 2020-05-29 株式会社岛津制作所 细胞评价用器件以及细胞评价用系统
CN111269833A (zh) * 2018-12-05 2020-06-12 中国科学院大连化学物理研究所 一种基于器官芯片的人胰岛类器官模型构建方法
CN111394248A (zh) * 2020-04-23 2020-07-10 西安交通大学口腔医院 一种细胞培养实验装置
JPWO2021132586A1 (fr) * 2019-12-27 2021-07-01
CN114317272A (zh) * 2021-12-07 2022-04-12 上海前瞻创新研究院有限公司 一种多细胞共培养的培养装置及细胞培养方法
US11976304B2 (en) 2016-09-13 2024-05-07 President And Fellows Of Harvard College Methods relating to intestinal organ-on-a-chip

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA3066338C (fr) * 2017-06-09 2023-03-14 Fujifilm Corporation Dispositif de micro-circuit fluidique

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH04278083A (ja) * 1991-02-28 1992-10-02 W R Grace & Co 細胞数を制御した細胞塊状体形成法およびそれに用いる細胞培養用基材
WO2005100549A1 (fr) * 2004-04-14 2005-10-27 Kanazawa University Technology Licensing Organization Ltd. Procédé de prolifération de cellules souches spécifiques d’un organe et appareil de prolifération pour celui-ci

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP4294425B2 (ja) * 2003-09-10 2009-07-15 独立行政法人農業・食品産業技術総合研究機構 細胞培養プレート
JP4456393B2 (ja) * 2004-03-26 2010-04-28 大日本印刷株式会社 細胞培養基板の製造方法および細胞培養基板製造装置
JP5281886B2 (ja) * 2006-04-28 2013-09-04 株式会社クラレ 細胞培養容器およびその製造方法
JPWO2009099066A1 (ja) * 2008-02-04 2011-05-26 株式会社島津製作所 バイオデバイスの製造方法

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH04278083A (ja) * 1991-02-28 1992-10-02 W R Grace & Co 細胞数を制御した細胞塊状体形成法およびそれに用いる細胞培養用基材
WO2005100549A1 (fr) * 2004-04-14 2005-10-27 Kanazawa University Technology Licensing Organization Ltd. Procédé de prolifération de cellules souches spécifiques d’un organe et appareil de prolifération pour celui-ci

Cited By (40)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11884938B2 (en) 2011-02-28 2024-01-30 President And Fellows Of Harvard College Cell culture system
US10655098B2 (en) 2011-02-28 2020-05-19 President And Fellows Of Harvard College Cell culture system
US10472612B2 (en) 2011-02-28 2019-11-12 President And Fellows Of Harvard College Cell culture system
EP2788469A1 (fr) * 2011-12-07 2014-10-15 A4TEC - Association for The Advancement of Tissue Engineering and Cell Based Technologies and Therapies Procédé à une seule étape et dispositif de génération de substituts de tissu tubulaire stratifié
JPWO2015159333A1 (ja) * 2014-04-14 2017-04-13 株式会社島津製作所 細胞培養用デバイス、細胞培養用システム及び細胞培養方法
US10301586B2 (en) 2014-04-14 2019-05-28 Shimadzu Corporation Cell culturing device, cell culturing system and cell culturing method
WO2015159333A1 (fr) * 2014-04-14 2015-10-22 株式会社島津製作所 Dispositif de culture cellulaire, système de culture cellulaire et procédé de culture cellulaire
JP2017514524A (ja) * 2014-05-08 2017-06-08 ウニヴェルジテーツクリニークム イェーナ 細胞層の配列のインビトロ産生方法およびデバイス
US20170226457A1 (en) * 2014-05-08 2017-08-10 Universitaetsklinikum Jena Method and Devices for the In Vitro Production of Arrangements of Cell Layers
US10731119B2 (en) 2014-05-08 2020-08-04 Universitaetsklinikum Jena Method and devices for the in vitro production of arrangements of cell layers
WO2015169287A1 (fr) * 2014-05-08 2015-11-12 Universitätsklinikum Jena Procédé et dispositifs pour la production in vitro d'ensembles de couches cellulaires
JP2016214148A (ja) * 2015-05-20 2016-12-22 住友電気工業株式会社 細胞培養担体及びこれを備える細胞シート
WO2017096192A1 (fr) 2015-12-04 2017-06-08 President And Fellows Of Harvard College Dispositifs de simulation d'une fonction d'un tissu hépatique et leurs procédés d'utilisation et de fabrication
GB2562396B (en) * 2015-12-04 2022-04-13 Harvard College Devices for simulating a function of a liver tissue and methods of use and manufacturing thereof
EP3383996A4 (fr) * 2015-12-04 2019-10-02 President and Fellows of Harvard College Dispositifs de simulation d'une fonction d'un tissu hépatique et leurs procédés d'utilisation et de fabrication
AU2016362491B2 (en) * 2015-12-04 2021-04-29 President And Fellows Of Harvard College Devices for simulating a function of a liver tissue and methods of use and manufacturing thereof
JP2019518443A (ja) * 2016-05-11 2019-07-04 ユニヴェルシテ グルノーブル アルプ 細胞共培養チップ及びその製造に関するプロセス
US11857958B2 (en) 2016-05-11 2024-01-02 Universite Grenoble Alpes Cell co-culture chip and process for the production thereof
JP6205507B1 (ja) * 2016-05-19 2017-09-27 光次 斉藤 培養装置および培養方法、並びにこの培養方法により製造された培養臓器
US12247227B2 (en) 2016-09-13 2025-03-11 President And Fellows Of Harvard College Methods relating to intestinal organ-on-a-chip
US12104174B2 (en) 2016-09-13 2024-10-01 President And Fellows Of Harvard College Methods relating to intestinal organ-on-a-chip
US11976304B2 (en) 2016-09-13 2024-05-07 President And Fellows Of Harvard College Methods relating to intestinal organ-on-a-chip
JP7011034B2 (ja) 2018-03-02 2022-02-10 富士フイルム株式会社 マイクロ流路デバイス
WO2019168118A1 (fr) * 2018-03-02 2019-09-06 富士フイルム株式会社 Dispositif à canaux microfluidiques
CN111788147A (zh) * 2018-03-02 2020-10-16 富士胶片株式会社 微流路装置
JPWO2019168118A1 (ja) * 2018-03-02 2021-01-14 富士フイルム株式会社 マイクロ流路デバイス
EP3744678A4 (fr) * 2018-03-02 2021-03-03 FUJIFILM Corporation Dispositif à canaux microfluidiques
JP2020010641A (ja) * 2018-07-18 2020-01-23 株式会社島津製作所 細胞培養装置および細胞播種方法
JP7158679B2 (ja) 2018-07-18 2022-10-24 株式会社島津製作所 細胞培養装置および細胞播種方法
JP2020080697A (ja) * 2018-11-21 2020-06-04 株式会社島津製作所 細胞評価用デバイス及び細胞評価用システム
JP7162836B2 (ja) 2018-11-21 2022-10-31 株式会社島津製作所 細胞評価用デバイス及び細胞評価用システム
US11549092B2 (en) 2018-11-21 2023-01-10 Shimadzu Corporation Cell evaluation device and cell evaluation system
CN111205959A (zh) * 2018-11-21 2020-05-29 株式会社岛津制作所 细胞评价用器件以及细胞评价用系统
CN111269833A (zh) * 2018-12-05 2020-06-12 中国科学院大连化学物理研究所 一种基于器官芯片的人胰岛类器官模型构建方法
JP7136509B2 (ja) 2019-12-27 2022-09-13 学校法人高崎健康福祉大学 肝細胞培養膜、それを備えた薬物輸送能評価キット、及び薬物輸送能評価方法
WO2021132586A1 (fr) * 2019-12-27 2021-07-01 学校法人高崎健康福祉大学 Membrane de culture de cellules hépatiques, kit d'évaluation de capacité de transport de médicament équipée de cette dernière, et procédé d'évaluation de capacité de transport de médicament
JPWO2021132586A1 (fr) * 2019-12-27 2021-07-01
CN111394248A (zh) * 2020-04-23 2020-07-10 西安交通大学口腔医院 一种细胞培养实验装置
CN114317272A (zh) * 2021-12-07 2022-04-12 上海前瞻创新研究院有限公司 一种多细胞共培养的培养装置及细胞培养方法
CN114317272B (zh) * 2021-12-07 2024-01-12 上海前瞻创新研究院有限公司 一种多细胞共培养的培养装置及细胞培养方法

Also Published As

Publication number Publication date
JPWO2012032646A1 (ja) 2013-12-12
JP5700460B2 (ja) 2015-04-15

Similar Documents

Publication Publication Date Title
JP5700460B2 (ja) 細胞培養デバイス及び細胞培養方法
Tehranirokh et al. Microfluidic devices for cell cultivation and proliferation
Ashammakhi et al. Kidney-on-a-chip: untapped opportunities
Xie et al. h-FIBER: Microfluidic topographical hollow fiber for studies of glomerular filtration barrier
Walker et al. Microenvironment design considerations for cellular scale studies
Caplin et al. Microfluidic organ‐on‐a‐chip technology for advancement of drug development and toxicology
Leclerc et al. Microfluidic PDMS (polydimethylsiloxane) bioreactor for large‐scale culture of hepatocytes
Inamdar et al. Microfluidic cell culture models for tissue engineering
JP6823792B2 (ja) 細胞培養装置および細胞培養方法
WO2009099066A1 (fr) Biodispositif
Zhang et al. Recent Advances in Microfluidic Platforms for Programming Cell‐Based Living Materials
EP3279310A1 (fr) Appareil et procédé de culture cellulaire
JP5686310B2 (ja) 細胞培養デバイス、細胞培養システム、及び細胞培養方法
US20080233607A1 (en) Cell Culture Device
Li et al. Selective stamp bonding of PDMS microfluidic devices to polymer substrates for biological applications
CN102257124A (zh) 芯片上器官装置
Marimuthu et al. Microfluidic cell coculture methods for understanding cell biology, analyzing bio/pharmaceuticals, and developing tissue constructs
WO2000078920A1 (fr) Procedes et dispositifs de culture cellulaire et systemes d'assitance d'organes
JP6823793B2 (ja) 細胞培養装置および細胞培養方法
CN112739812A (zh) 堆叠的再循环生物反应器
CN113174332A (zh) 仿生肾器官芯片结构和仿生肝肾芯片结构
CN110257243B (zh) 微流控芯片打印喷嘴和生物3d打印系统
Khademhosseini Micro and nanoengineering of the cell microenvironment: technologies and applications
WO2011023655A1 (fr) Système microfluidique et son procédé de production
JP5731421B2 (ja) 細胞培養デバイス

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 10856992

Country of ref document: EP

Kind code of ref document: A1

ENP Entry into the national phase

Ref document number: 2012532811

Country of ref document: JP

Kind code of ref document: A

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 10856992

Country of ref document: EP

Kind code of ref document: A1

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载