Method for maturation and amplification of hepatocyte-like cells

A technology of liver-like cells and hepatocytes, which is applied in the field of biomedicine and cell transformation, can solve the problems of limited number of hepatocytes, high operation requirements, and high cost, and achieve the solution of immature hepatocytes, large amount of injected cells, and rapid expansion Effect

Pending Publication Date: 2020-04-28
立沃生物科技(深圳)有限公司
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Problems solved by technology

However, the source of hepatocytes has restricted its wide application
The liver cells obtained by the method of in vitro regeneration of liver cells are liver-like cells, express some liver cell marker genes, and only have part of the physiological functions of liver cells, which is still far behind the primary human liver cells
In addition, the method of in vitro

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  • Method for maturation and amplification of hepatocyte-like cells
  • Method for maturation and amplification of hepatocyte-like cells
  • Method for maturation and amplification of hepatocyte-like cells

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Embodiment Construction

[0038] The realization of the purpose of the present invention, functional characteristics and advantages will be further described in conjunction with the embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0039] The invention provides a method for maturation and expansion of hepatic cells, the specific steps comprising:

[0040] Step S1: the stem cells are cultured in vitro using a corresponding induction medium to induce stem cells to differentiate into liver-like cells;

[0041] Step S2: Inject the liver-like cells into the liver carrier of the FAH gene-deficient rat through the spleen;

[0042] Step S3: Stimulating the apoptosis of the autologous hepatocytes of the FAH gene-deficient rat liver carrier, and promoting the maturation and proliferation of hepatoid cells;

[0043] Step S4: Separation and purification to obtain primary human hepatocytes.

[00...

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Abstract

The invention relates to a method for maturation and amplification of hepatocyte-like cells. The method comprises the following steps: S1, culturing stem cells in vitro by using an induction liquid toinduce mesenchymal stem cells to differentiate into hepatocyte-like cells; S2, injecting the hepatocyte-like cells into the liver vector of an FAH gene-deficient rat through the spleen in a fixed-point manner; S3, stimulating FAH gene-deficient rat liver vector autologous hepatocyte apoptosis, and promoting maturation and proliferation of the hepatocyte-like cells; and S4, separating and purifying to obtain human primary hepatocyte. Human mesenchymal stem cells are induced in vitro to form the hepatocyte-like cells, the hepatocyte-like cells are injected into the liver of the FAH gene-deficient rat in the fixed-point manner through the spleen, and the internal environment of the liver of the FAH gene-deficient rat is utilized to efficiently differentiate the hepatocyte-like cells into mature human hepatocytes and realize rapid amplification of the hepatocytes. Therefore, the defects of immaturity, small scale, high cost and the like of the hepatocytes obtained by a hepatocyte in-vitroregeneration method are overcome.

Description

【Technical field】 [0001] The invention relates to the technical field of biomedicine, to the technical field of cell transformation, and in particular to a method for maturation and expansion of hepatic cells. 【Background technique】 [0002] Primary human hepatocytes refer to hepatocytes cultured immediately after removal from human liver tissue. They can be used in basic life science research, drug development, liver cell transplantation, and 3D printing of liver organs. The market capacity can reach 100 billion. However, the source of hepatocytes has restricted its wide application. On the one hand, due to cultural factors, the organ donation rate of Chinese citizens after death is extremely low; on the other hand, my country's laws and regulations on organ donation are not perfect, which leads to the extreme shortage of primary human liver cells. [0003] At present, the technical means to solve the source of primary human hepatocytes mainly include: 1) Liver cell regene...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N5/0775C12N5/074C12N5/0735
CPCC12N5/067C12N2506/02C12N2506/1353C12N2506/45C12N2501/237C12N2501/16C12N2500/32C12N2501/415C12N2501/155C12N2501/119C12N2501/12C12N2501/39C12N2500/44C12N2500/30C12N2500/25C12N2501/115C12N2501/11
Inventor 周明
Owner 立沃生物科技(深圳)有限公司
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