Method for maturation and amplification of hepatocyte-like cells

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...

AI Technical Summary

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 regeneration of liver cells has the disadvantages of high operational requirements, small scale, and high cost, and is not feasible in practical applications.

The method of in vivo expansion of hepatocytes (mice) can obtain a certain number of mature primary human hepatocytes, but cannot achieve personalized customization, and the number of hepatocytes is limited

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