Sodium butyrate induces mouse embryonic stem cells to differentiate into hepatocytes in vitro / 中国病理生理杂志

ABSTRACT

AIM: To explore mechanism by which sodium butyrate induces mouse embryonic stem cells(ES) to differentiate into hepatocytes in vitro.METHODS: E14 mouse ES cells were cultivated in a routine way,and then cultivated in suspension to form embryonic bodies(EBs).EBs were transferred into 6-well culture dishes and 3 mmol/L sodium butyrate was added into the culture medium.Morphological changes were investigated by phase contrast microscopy.?-fetoprotein(AFP),albumin(ALB) and cytokeratin 18(CK18) were examined by immunofluorescence staining.AFP,ALB,?_1-antitrypsin(AAT) and TTR mRNA were assayed by RT-PCR.Proportion of ALB positive cells was analyzed by flow cytometry.Periodic acid Schiff(PAS) reaction and indocyanine green(ICG) uptake assay were performed to assess the characteristic hepatocyte function of the differentiated cells.RESULTS: In the presence of sodium butyrate,parts of ES cells differentiated into a population with epithelial morphology similar to mouse hepatocytes.AFP and TTR mRNA expression were observed at 7 d,and ALB and AAT mRNA expressed at 14 d.Hepatocytes specific markers,ALB,AFP and CK18 were positive expression in immunofluorescence staining at 14 d.PAS reaction and ICG uptake were positive for the hepatocyte-like cells.CONCLUSION: Mouse ES cells can be induced into hepatocyte-like cells by sodium butyrate efficiently,and these ES cells-derived hepatocytes possess characteristic hepatocytic function.