The Effect Of Astragalus Injection On The Differentiation Of Human Parthenogenetic Embryonic Stem Cells Into Hepatocyte-like Cells

Background and Objectives: Acute-on-chronic liver failure(ACLF)is a critical disease in clinical practice and has a high mortality rate.There is still a lack of ideal treatment for ACLF.Adopting stem cell transplantation to treat ACLF has become a new research direction.Human parthenogenetic embryonic stem cells(h PESCs)not only have the same ability to differentiate as human embryonic stem cells(hESCs),but also avoid ethical controversies and transplantation immune rejection.Besides,Pharmacological studies also show that astragalus has protective effects on experimental hepatitis,and avoid degeneration and necrosis of liver cells,regulate immunity,and promote hepatocyte-like cells regeneration through multiple links.The aim of this study was to explore the effects of chemical inducers on the differentiation of hPESCs into hepatocyte cells by adding astragalus injection.Methods: 1)Establish hPESCs/3T3 culture system in vitro.The growth state of hPESCs was observed.Morphological observation and alkaline phosphatase(AKP)were used to determine whether hPESCs could grow steadily and remain undifferentiated in the culture system.2)hPESCs was induced to hepatocytes-like cells by chemical inducers and astragalus injection plus chemical inducers.The expression of Sox17,Gsc alpha fetoprotein(AFP),albumin(ALB),CK18 and Hepa to determine whether the induced cells have hepatocyte function.Results: 1)hPESCs grew well in hPESCs/3T3 culture system.After more than 30 generations of in vitro culture,hPESCs still maintained its biological characteristics.2)The cell morphology of Astragalus plus chemical induction group and chemically induced group h PESCs was similar to human hepatocytes after induction.3)The expressions of Sox17,Gsc,AFP and ALB in chemical inducible group and Astragalus plus chemical induction group were significantly higher than those in 0d.The expression of each index in chemical induction group was higher than that inAstragalus plus chemical induction group.4)CK18 and Hepa protein were positive in two experimental groups.5)The contents of AFP,ALB and BUN in the two experimental groups were significantly higher than those in the control group,and the contents in the chemical induction group were all higher than those in the Astragalus plus chemical induction group.Conclusions: 1)In the hPESCs/3T3 culture system,hPESCs can effectively proliferate and maintain its undifferentiated state.2)Two experimental groups can inhibit the differentiation of hPESCs into hepatocyte-like cells with certain functions.3)Astragalus injection has inhibitory effect on differentiation of hPESCs into-like cells.