Recoverable immortalized hepatic cell line carrying double suicide genes and establishing method of recoverable immortalized hepatic cell line

Abstract

The invention provides an establishing method of a recoverable immortalized hepatic cell line. The establishing method comprises the following steps: separating a hepatic progenitor cell from liver in a mouse which is 12.5-14.5 days old in an embryonic period; guiding a retrovirus containing genes SV40T and HSV-TK into the hepatic progenitor cell; screening a monoclonal cell strain having a hepatic progenitor cell marker and having a function of differing to a mature hepatic cell; then guiding a retrovirus containing CD gene into the cell strain so as to obtain recoverable immortalized hepatic cell carrying double suicide genes. The cell strain can multiply in vitro to obtain the phenotype and functions of a normal hepatic cell, the safety of the immortalized cell becomes adjustable through controllable modes such as locus recombination and drug screening; when the cell is used in a human body, the biological safety of the immortalized cell is ensured to the greatest extent and the dangerousness of the immortalized cell is reduced; therefore, a reliable, safe and ideal hepatic cell material is provided for bioartificial liver technology.

Description

technical field [0001] The invention relates to the field of cell biology, and discloses a construction method of an immortalized liver cell strain carrying double suicide genes and a constructed mouse embryonic liver progenitor cell strain. technical background [0002] Liver failure is the most common clinical syndrome of liver diseases with extremely high mortality, which seriously threatens human health. According to statistics, 300,000 to 500,000 people die each year due to severe liver failure in my country. The most effective treatment for liver failure is liver transplantation. However, due to the lack of donor liver sources, less than 10% of patients can receive liver transplantation. Bioartificial liver (BAL) is an in vitro bioartificial organ device based on culturing hepatocytes, which can temporarily replace the liver to perform functions such as detoxification, secretion, synthesis, and transformation. It can not only buy time for liver failure patients waitin...

AI Technical Summary

Problems solved by technology

In the Cre-LoxP protocol, the activity of Cre recombinase and the efficiency of recombination and excision of the LoxP site cannot be controlled, and it cannot be guaranteed that all cells will remove the immortalized gene. After the restoration of immortalization, whether the remaining immortalized gene in the cell inhibits the function of the cell? There are problems such as the risk of tumorigenesis and cell transformation that need to be further verified

Although the suicide gene program increases the safety of cells in vivo, the immortalized cells introduced into the body still have the SV40T gene, which affects the differentiation and functional maturation of cells in vivo

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