WO2017126758A1 - Unité de culture d'hépatocytes en trois dimensions, système d'évaluation d'hépatotoxicité et procédé d'évaluation de l'hépatotoxicité utilisant celui-ci - Google Patents

Unité de culture d'hépatocytes en trois dimensions, système d'évaluation d'hépatotoxicité et procédé d'évaluation de l'hépatotoxicité utilisant celui-ci Download PDF

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WO2017126758A1
WO2017126758A1 PCT/KR2016/008549 KR2016008549W WO2017126758A1 WO 2017126758 A1 WO2017126758 A1 WO 2017126758A1 KR 2016008549 W KR2016008549 W KR 2016008549W WO 2017126758 A1 WO2017126758 A1 WO 2017126758A1
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culture
fluid
liver
constituent cells
dimensional
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PCT/KR2016/008549
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Korean (ko)
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윤석주
오정화
안재환
김우근
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한국화학연구원
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C12M25/14—Scaffolds; Matrices
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing

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  • the present invention relates to a three-dimensional hepatocyte culture unit, hepatotoxicity evaluation system and hepatotoxicity evaluation method using the same.
  • the liver is a major living organ of an animal, and the main function of the liver is a detoxification function that detoxifies foreign substances.
  • the liver is composed of unit structures called hepatic lobules, and these hepatic lobules may have a substantially hexagonal structure.
  • hepatocytes extracted from the human body can be cultured for a long time using a microstructure that simulates the boundary between epithelial tissue and blood vessels of the liver and expresses a function of liver tissue such as albumin production to form a structure similar to the bile duct in the human body.
  • Research has been reported to develop liver chips. (See An Artificial Liver Sinusoid With a Microfluidic Endothelial-Like Barrier for Primary Hepatocyte Culture, Lee et al. Biotechnology and Bioengineering, 97, 5, 1312, 2007).
  • Embodiments of the present invention co-cultivate various cells constituting the liver tissue by simulating the structure and environment of hepatic lobule, which is a unit structure of the liver, and using the 3D hepatocyte culture unit, hepatotoxicity evaluation system, and hepatotoxicity using the same for evaluating hepatotoxicity An evaluation method is provided.
  • the fluid including at least one of the may include a second culture structure that can be accommodated therein.
  • the second culture structure may include at least one first injection means into which the fluid is injected, at least one first groove portion into which one side of the first culture structure is inserted, and the other side of the first culture structure. It may include at least one second groove portion, and may include a first drain means for draining at least a portion of the fluid injected from the first injection means.
  • first drainage means may be provided to discharge the fluid contained in the second culture structure by lowering from the top to the bottom of the second culture structure.
  • the apparatus may further include a receiving case accommodating the second culture structure while closing one end of the first injection means and one end of the first drain means.
  • the first culture structure may include a porous matrix that provides a culture space in which the liver constituent cells are cultured.
  • the liver constituent cells may include at least one of hepatocytes, kupffer cells and stellate cells.
  • the first culture structure and the at least one first culture structure in which the liver constituent cells are cultured is inserted, the culture medium for culturing the liver constituent cells and hepatotoxic substance for hepatotoxicity evaluation
  • a hepatotoxicity evaluation system can be provided that includes a fluid supply member for supplying a fluid to a three-dimensional hepatocyte culture unit.
  • the plate is formed on the flow path, the flow path, the fluid is moved, at least one or more second injection means and the second injection means for injecting the fluid from the flow path to the three-dimensional hepatocyte culture unit And second drainage means for draining at least some of the fluid injected therefrom.
  • one end of the flow path may be connected to the fluid supply member, and the branch point of the flow path may be a point extending from one end of the flow path connected to the fluid supply member.
  • the apparatus may further include a fluid discharge member positioned on the other side of the plate to discharge the fluid to the outside.
  • the step of providing a three-dimensional hepatocyte culture unit having a second culture structure is inserted into the first culture structure in which the liver constituent cells are cultured, accommodating the three-dimensional hepatocyte culture unit in a housing case Supplying a fluid including at least one of a culture medium for culturing the liver constituent cells and a hepatotoxic substance for hepatotoxicity evaluation, to the three-dimensional hepatocyte culture unit, and the fluid from the three-dimensional hepatocyte culture unit. Discharging the first culture construct from the second culture construct to measure the degree of necrosis of the liver constituent cells and analyzing cytokines generated in the fluid discharged to the outside. Hepatotoxicity assessment methods may be provided.
  • a three-dimensional hepatocyte culture unit having a second culture structure in which a first culture structure in which liver constituent cells are cultured is inserted, wherein the three-dimensional hepatocyte culture unit is at least placed on a plate.
  • the providing of the three-dimensional hepatocyte culture unit may include preparing the first culture structure, culturing the liver constituent cells in the first culture structure, and applying the first culture structure to the second culture structure. It may include inserting into the installation.
  • the step of preparing the first culture structure May be performed through at least one process.
  • the step of culturing the liver constituent cells in the first culture structure may include the step of stabilizing.
  • the method may further include culturing the liver constituent cells cultured in the first culture structure after inserting the first culture structure into the second culture structure.
  • the fluid supplied by the fluid supply member is automatically discharged to the outside according to the flow of the fluid flowing from one side of the plate to the other side of the plate, or the outside by the fluid discharge member located on the other side of the plate Can be discharged.
  • Embodiments according to the present invention co-culture the liver constituent cells in a three-dimensional hepatocyte culture unit similar to the structure and environment of hepatic lobule, a liver unit structure, and hepatotoxicity evaluation in a hepatotoxicity evaluation system employing at least one three-dimensional hepatocyte culture unit Since the continually proceeds, hepatotoxicity evaluation reflecting the mutual linkage between the liver constituent cells is possible.
  • FIG. 1 is a perspective view showing a three-dimensional hepatocyte culture unit according to an embodiment of the present invention.
  • FIG. 2 is a perspective view showing the first culture structure of FIG. 1.
  • FIG. 3 is a perspective view illustrating the second culture structure of FIG. 1.
  • FIG. 4 is a perspective view showing a combined state of the three-dimensional hepatocyte culture unit of Figure 1 accommodated in the housing case.
  • FIG. 5 is a perspective view showing a hepatotoxicity evaluation system in which the three-dimensional hepatocyte culture unit of FIG. 1 is installed.
  • FIG. 6 is a perspective view showing the flow of the fluid in the hepatotoxicity evaluation system of FIG.
  • FIG. 1 is a perspective view showing a three-dimensional hepatocyte culture unit according to an embodiment of the present invention
  • Figure 2 is a perspective view showing a first culture structure of Figure 1
  • Figure 3 is a perspective view showing a second culture structure of Figure 1 .
  • the 3D hepatocyte culture unit 10 may include a first culture structure 100 and a second culture structure 200.
  • At least one first culture structure 100 may be provided and may be inserted into the second culture structure 200.
  • the number of the first culture structures 100 inserted into the second culture structure 200 may vary depending on the purpose and / or conditions of the user.
  • the first culture structure 100 may have a porous structure, for example, the first culture structure 100 may be a lattice structure.
  • the lattice may have the shape of a groove or the shape of a hole.
  • the lattice may be freely applied as the shape of the lattice if the liver constituent cells have a predetermined space to be cultured.
  • the first culture structure 100 may be implemented in various forms such as a sponge structure, a hydrogel, a solution in which a biopolymer is dissolved in a liquid medium.
  • the first culture structure 100 may be composed of, for example, polycaprolactone (PCL), polylactic acid (polylactic acid, PLA) and the like. However, this is only an example, and if the material is biocompatible and has a hardness of a certain level or more, it may be applied as a material constituting the first culture structure 100.
  • PCL polycaprolactone
  • PLA polylactic acid
  • liver constituent cells including at least one of hepatocytes, cooper cells, and stellate cells may be cultured.
  • the first culture structure 100 may be inserted into the second culture structure 200 in a state that the liver constituent cells are cultured for each lattice.
  • the liver is inserted into the first culture structure 100 while the first culture structure 100 is inserted into the second culture structure 200.
  • the constituent cells can also be cultured.
  • the second culture structure 200 may be inserted into the first culture structure 100, and may have a receiving space for accommodating a fluid such as culture medium, hepatotoxic substances, etc., the receiving space is an example of a cylindrical shape Can be formed.
  • the second culture structure 200 may include a first injection means 210, a first recess 220, and a first drain means 230.
  • At least one first injection means 210 may be provided at the bottom of the second culture structure 200.
  • the first injection means 210 may be spaced apart at predetermined intervals along the circumference of the bottom of the second culture structure 200.
  • first injection means 210 may be provided along the circumference of the bottom of the second culture structure 200, and three first injection means 210 may be provided with a second drainage means 230. It may be formed at equal intervals on the circumference of the imaginary circle with the center.
  • the first injection means 210 may be formed in the form of a hole.
  • fluid such as culture medium, hepatotoxic substances, etc. may be injected into the second culture structure 200 through the first injection means 210.
  • At least one first recess 220 may be provided on a circumferential wall of the second culture structure 200, and one side of the first culture structure 100 may be inserted into the first recess 220. have.
  • the first drainage means 230 may be provided in the central portion of the second culture structure 200, the other side of the first culture structure 100 is inserted, and substantially the number of the first recesses 220.
  • the second recess 225 having the same number may be provided.
  • the first drain means 230 may be formed in a cylindrical shape as an example.
  • the first culture structure 100 since the first culture structure 100 is fitted into the first groove portion 220 and the second groove portion 225, the first culture structure 100 can be selectively inserted into the second culture structure 200. Since the number of the first culture structures 100 to be inserted into the two culture structures 200 can be freely changed, hepatocyte culture efficiency and / or hepatotoxicity evaluation efficiency can be increased.
  • the first culture structure 100 is described as being fitted by the first groove portion 220 and the second groove portion 225, the first culture structure 100 and the second culture structure.
  • the connection relationship of the 200 is not limited thereto.
  • the first culture structure 100 may be fixed by controlling magnetism, pressure, and the like between the second culture structures 200.
  • the first drain means 230 may drain at least a portion of the fluid injected through the first injection means 210.
  • the first drainage means 230 may be provided to allow the fluid contained in the second culture structure 200 to be discharged by moving from the top to the bottom of the second culture structure 200.
  • the height of the first drainage means 230 may be formed to be relatively lower than the height of the second culture structure (200). Accordingly, even when the fluid is continuously injected into the second culture structure 200 through the first injection means 210, the fluid may be drained in advance by the first drain means 230 having a relatively low height. Therefore, the fluid can be prevented from overflowing from the second culture structure 200 to the outside.
  • the height of the first drainage means 230 may be the same as the height of the second culture structure 200.
  • the first recess 220 and the second recess 225 are each provided with twelve, but the number of the first recess 220 and the second recess 225 is limited thereto.
  • the number of the first recesses 220 and the second recesses 225 may vary depending on the number of the first culture structures 100 inserted into the second culture structures 200.
  • the three-dimensional hepatocyte culture unit 10 is a liver constituent cell in the second cell culture structure 200 which is a structure that simulates the structure and environment of hepatic lobule which is the basic structure of the liver.
  • the cultured first culture structure 100 may be selectively inserted and used for evaluating hepatotoxicity.
  • the hepatotoxicity evaluation may be performed using liver constituent cells cultured in the first culture construct 100, or the first culture construct 100 in which the liver constituent cells are cultured may be stored in the second culture construct 200. Insertion may be performed using additionally cultured liver constituent cells.
  • FIG. 4 is a perspective view showing a combined state of the three-dimensional hepatocyte culture unit of Figure 1 accommodated in the housing case.
  • the 3D hepatocyte culture unit 10 may be accommodated in the accommodation case 15.
  • the accommodating case 15 may have a hexagonal shape and may further include an accommodating space having a cylindrical shape therein to accommodate the three-dimensional hepatocyte culture unit 10.
  • the first injection means 210 of the second culture structure 200 is accommodated by the storage case 15.
  • One end of) and one end of the first drainage means 230 may be closed. Accordingly, the loss of fluid that may occur due to the first injection means 210 and the first drain means 230 can be minimized.
  • the housing case 15 may be a member required when the 3D hepatocyte culture unit 10 is to be used alone.
  • the three-dimensional hepatocyte culture unit 10 may be accommodated in the accommodating case 15 and used alone, as described above.
  • the three-dimensional hepatocyte culture unit 10 includes at least one three-dimensional hepatocyte culture unit 10. It is also possible to be used as the hepatotoxicity evaluation system 1000.
  • the detailed configuration of the hepatotoxicity evaluation system 1000 will be described with reference to FIGS. 5 and 6.
  • FIG. 5 is a perspective view showing a hepatotoxicity evaluation system in which the three-dimensional hepatocyte culture unit of FIG. 1 is installed
  • FIG. 6 is a perspective view showing the flow of fluid in the hepatotoxicity evaluation system of FIG. 5.
  • the hepatotoxicity evaluation system 1000 may include at least one three-dimensional hepatocyte culture unit 10.
  • the hepatotoxicity evaluation system 1000 may include a three-dimensional hepatocyte culture unit 10, a plate 300, and a fluid supply member 400.
  • the hepatotoxicity evaluation system 1000 may further include a fluid discharge member 500.
  • the three-dimensional hepatocyte culture unit 10 includes a first culture structure 100, a second culture structure 200 including a first injection means 210, a first recess 220, and a first drainage means 230. It may include. However, since the three-dimensional hepatocyte culture unit 10 has been described with reference to FIGS. 1 to 4, the overlapping description of the previously described portions will be omitted.
  • At least one three-dimensional hepatocyte culture unit 10 may be installed on the plate 300, and may include a flow path 310, a second injection means 320, and a second drain means 330.
  • the flow path 310 may be formed inside the plate 300, and as shown by a large arrow in FIG. 6, the constituent cells are cultured while the fluid including the culture medium and the hepatotoxic substance flows along the flow path 310. Or hepatotoxicity assessment may be made. However, this is only an example, and the flow path 310 may be formed on the bottom surface of the plate 300.
  • the bottom surface of the plate 300 means a surface in which the three-dimensional hepatocyte culture unit 10 is in contact with the plate 300.
  • the flow path 310 may be formed by segmenting the number of three-dimensional hepatocyte culture unit 10 installed in the hepatotoxicity evaluation system 1000, the second drainage means 330 on the upstream side and the downstream side adjacent thereto. It may include a connection passage 312 for connecting the second injection means 320 of the.
  • a branch point 311 may be formed at the end of the second injection means 320 side of the connection path 312 included in the flow path 310, and the flow path 310 is at least two branches by the branch point 311. Can be cracked.
  • the flow path 310 is formed at the periphery of the three-dimensional hepatocyte culture unit 10 when viewed from the center of one three-dimensional hepatocyte culture unit 10. It may extend along at least a portion.
  • second injection means 320 may be formed at each end of the branch point 311 and the flow path 310 branching from the branch point 311.
  • the second injection means 320 may be formed to protrude vertically from the plate 300, the first injection of the three-dimensional hepatocyte culture unit 10 when the three-dimensional hepatocyte culture unit 10 is installed in the plate 300 It may be formed at a position that can penetrate the means 210. Accordingly, the fluid flowing through the flow passage 310 rises to the top of the three-dimensional hepatocyte culture unit 10 by the second injection means 320, and the risen fluid is from the top of the three-dimensional hepatocyte culture unit 10. It can spread inside.
  • the second injection means 320 may be provided in the form of microtubules, for example, and may be configured to allow the fluid to rise by capillary action.
  • the second drainage means 330 may be provided to be fitted to the first drainage means 230 formed in the second culture structure 200 of the three-dimensional hepatocyte culture unit 10. Accordingly, the diameter of the second drain means 330 may be formed to be substantially smaller than the diameter of the first drain means 230.
  • This second drainage means 330 is an amount exceeded if the amount of the fluid injected from the second injection means 320 exceeds the amount of fluid that the three-dimensional hepatocyte culture unit 10 can accommodate. The fluid may be lowered from the top to the bottom of the three-dimensional hepatocyte culture unit 10 to drain the fluid.
  • the fluid supply member 400 may be located at one side of the plate 300 and may be, for example, a fluid supply motor capable of continuously supplying fluid to the three-dimensional hepatocyte culture unit 10 installed at one end of the plate 300. Can be. In addition, the fluid supply member 400 may be connected to one end of the flow path 310 to continuously inject the fluid into the three-dimensional hepatocyte culture unit 10.
  • the fluid supplied from the fluid supply member 400 may be automatically discharged to the outside according to the flow of the fluid flowing from one side of the plate 300 to the other side of the plate 300, the fluid discharged to the outside Can be used to assess hepatotoxicity.
  • the method of discharging the fluid to the outside is not limited thereto.
  • the fluid may be discharged to the outside by the fluid discharge member 500.
  • the fluid discharge member 500 may include a plate. It may be located on the other side of the 300, for example, may be a fluid discharge motor capable of discharging the fluid from the three-dimensional hepatocyte culture unit 10 installed on the other end of the plate (300).
  • the fluid discharge member 500 may be connected to the other end of the flow path 310 to continuously discharge the fluid from the three-dimensional hepatocyte culture unit 10.
  • the hepatotoxicity evaluation system 1000 may further include a sensor and a controller.
  • the sensor may be provided in the vicinity of the second drainage means 330 of the three-dimensional hepatocyte culture unit 10 to sense the amount of the fluid continuously injected into each three-dimensional hepatocyte culture unit 10.
  • a flow sensor, a level sensor, or the like may be used.
  • the installation position and type of the sensor are not limited thereto.
  • control unit pre-stores data about the amount of fluid that can be accommodated in the three-dimensional hepatocyte culture unit 10, and compares the amount of the fluid sensed by the sensor with the amount of the pre-stored fluid, On the basis of this, the fluid injection speed of the fluid supply member 400 and the fluid discharge speed of the fluid discharge member 500 may be controlled. Accordingly, it is possible to prevent the inundation of the fluid which may occur in the process of continuously supplying the fluid to the three-dimensional hepatocyte culture unit 10, and to control the amount of the fluid more accurately, the hepatotoxicity evaluation system 1000 ) Can be operated.
  • the hepatotoxicity evaluation system 1000 since the hepatotoxicity evaluation system 1000 according to an embodiment of the present invention continuously injects and exchanges a culture medium into a plurality of three-dimensional hepatocyte culture units 10 connected to each other, an oxygen permeation amount is increased. It can improve the function of liver constituent cells. Hepatotoxicity can then be injected into the cultured liver constituent cells to continue the hepatotoxicity assessment.
  • the hepatotoxicity evaluation is performed by the hepatotoxicity evaluation system 1000 including the plurality of three-dimensional hepatocyte culture units 10, but this is only an example.
  • the three-dimensional hepatocyte culture unit 10 may be accommodated in the accommodating case 15, and then used alone to evaluate hepatotoxicity.
  • liver toxicity evaluation method will be described with reference to Examples.
  • Example 1 3D Hepatocyte culture and Hepatotoxicity evaluation
  • a first culture structure was prepared in which the physical properties of the pores were controlled.
  • the three-dimensional printing process used a nozzle having a diameter of about 0.2 mm, about 170 Under a temperature of o C to about 270 o C, a printing speed of about 30 mm s ⁇ 1 to 120 mm s ⁇ 1 was performed.
  • PCL polycaprolactone
  • PLA polylactic acid
  • the first culture construct was designed using 3ds Max Design (Autodesk, Inc., San Rafael, Calif., USA) with a lattice structure of about 0.1 mm to about 0.3 mm in diameter.
  • the second culture structure was a cylindrical structure having a diameter of about 4cm to about 8cm.
  • the solution was dissolved in a liquid medium and the liver constituent cells were agitated for 24 hours or more, thereby increasing the cell settling rate in the first culture construct. .
  • liver constituent cells human primary hepatoctyes, HepaRG or liver cancer cell lines (HepG2, Hep3B, Huh7) and the like can be utilized.
  • Cooper cells can be utilized as a human kupffer cell or established cell line U937.
  • LX2 cell line which is an established cell line can be utilized.
  • the first culture construct in which the liver constituent cells are primaryly cultured is applied to the second culture construct in an appropriate ratio (hepatocytes: Cooper cells, 3: 1 or 4: 1, hepatocytes: astrocytes, 3: 1 or 4: 1). After insertion, the culture medium was co-cultured by filling the height of the second culture construct.
  • acetoaminophen and triglitazone were injected into the second culture structure (hereinafter, 3-dimensional hepatocyte culture unit) into which the first culture structure was inserted at an appropriate concentration. Subsequently, after exposing the three-dimensional hepatocyte culture unit for a predetermined time, the first culture construct was separated from the second culture construct. Subsequently, the first culture construct exposed to the hepatotoxic substance was subjected to the cell counting kit-8 (Dojindo Laboratory, Kumanoto, Japan) or The celltiter-glo® luminescent cell viability assay (Promega Corp, Madison, USA). Cell viability in the first culture construct was measured. In addition, the degree of cell necrosis was measured by measuring lactate dehydrogenase (LDH), a cell damage index.
  • LDH lactate dehydrogenase
  • fluid eg, culture medium exposed for a period of time after injection of hepatotoxic material
  • fluid eg, culture medium exposed for a period of time after injection of hepatotoxic material
  • Cytokines cytokines
  • interleukin2 interleukin2
  • interlekin6 interleukin8
  • the method for evaluating hepatotoxicity comprises co-culturing liver constituent cells in a three-dimensional hepatocyte culture unit similar to the structure and environment of hepatic lobule, which is a unit structure of the liver, and Since hepatotoxicity evaluation is continuously performed in the hepatotoxicity evaluation system employing at least one or more, hepatotoxicity evaluation reflecting the mutual link between the liver constituent cells is possible.
  • first culture structure 200 second culture structure
  • first injection means 220 first groove portion
  • junction 312 connection

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Abstract

L'invention concerne une unité de culture d'hépatocytes en trois dimensions. L'unité de culture d'hépatocytes en trois dimensions selon un mode de réalisation de la présente invention comprend une première structure de culture dans laquelle sont cultivées des cellules formant le foie, et une deuxième structure de culture dans laquelle peut être insérée la première structure de culture et qui peut accueillir un fluide comprenant au moins un parmi un milieu de culture destiné à la culture des cellules formant du foie et un matériau hépatotoxiques destiné à évaluer l'hépatotoxicité.
PCT/KR2016/008549 2016-01-21 2016-08-03 Unité de culture d'hépatocytes en trois dimensions, système d'évaluation d'hépatotoxicité et procédé d'évaluation de l'hépatotoxicité utilisant celui-ci WO2017126758A1 (fr)

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WO2019059702A3 (fr) * 2017-09-25 2019-08-08 아주대학교산학협력단 Système de culture tridimensionnelle à long terme d'hépatocytes primaires basé sur des nanofibres et procédé de culture associé
CN112063525A (zh) * 2019-06-10 2020-12-11 芬欧汇川集团 细胞培养板，其制备方法和检测物质的方法
CN114080316A (zh) * 2019-07-23 2022-02-22 T&R碧欧法博有限公司 肝脏类器官及其制备方法
US11685900B2 (en) 2017-09-25 2023-06-27 NANOFAENTECH CO., Ltd. Nanofiber-based long-term primary hepatocyte three-dimensional culture system and culturing method

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KR102015562B1 (ko) 2017-12-04 2019-08-28 한국화학연구원 극성을 갖는 간세포 유사세포로의 분화를 유도하는 조성물 및 상기 조성물을 이용한 간세포 유사세포로의 분화 방법
US11732232B2 (en) 2020-08-14 2023-08-22 Korea Research Institute Of Bioscience And Biotechnology Biomimetic cell culture apparatus and cell culture system comprising the same

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