Method for inducing hepatic oval cells differentiation to bile duct cells and its special culture medium

Abstract

The invention discloses a method for inducing hepatic oval cells to differentiate towards bile duct cells and a special culture medium used in the method. The culture medium is added with 1 to 6 percent of fetal calf serum, 1 to 3 percent of B27 and 0.05 to 0.15 percent of penicillin-streptomycin on the basis of an RPMI1640 culture medium. The method for inducing the differentiation comprises the following steps: 1) leading RhoA mutant genes into the oval cells WB-F344 to obtain WB-F344 cells of a high-expression RhoA gene activity mutant; and 2) inoculating the WB-F344 cells of the high-expression RhoA gene activity mutant into the special culture medium where the hepatic oval cells are induced in vitro to differentiate into the bile duct cells, and culturing the WB-F344 cells at a temperature of 37 DEG C with 5 percent of CO2 to obtain the bile duct cells. The method provides a new approach to obtain the bile duct cells and has wide application prospect.

Description

technical field [0001] The invention relates to a method for inducing cell differentiation and its special medium, in particular to a method for inducing the differentiation of hepatic oval cells into bile duct cells and its special medium. Background technique [0002] WB-F344 cells are a type of cells isolated from the liver of Fischer male mice, which have many characteristics of hepatic stem / progenitor cells: 1) it has the phenotypic characteristics of hepatocytes and hepatic cholangiocytes; 2) when it is transplanted into rats 3) WB-F344 cells can develop into cholangiocarcinoma after chemical mutagenesis and transplanted into rat liver, indicating that it has the potential to differentiate into cholangiocytes, These characteristics all indicate that WB-F344 cells are hepatic stem / progenitor cells. Studies have found that WB-F344 cells treated with sodium butyrate have a series of markers of cholangiocytes, indicating that WB-F344 can differentiate into cholangiocytes ...

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Problems solved by technology

Li Wei et al. used RhoGDI expression to inhibit the activation of RhoA during the development of mouse embryos, and found that the inactivation of RhoA would lead to the death of mouse embryos at 7 days of development (Wei L, Imanaka-Yoshida K, Wang L, et al .Inhibition of Rho family GTPases by Rho GDPdissociation inhibitor disrupts cardiac morphogenesis and inhibits cardiomyocyte proliferation.Development, 2002,129(7):1705~1714); Decrease in embryo size and even death of the embryo

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