Induction of Rhesus Monkey Embryonic Stem Cells into Hepatocyte-like Cells by a Three-step Method

Embryonic stem cells (ESCs) hold great potential for the treatment of liver diseases. Here, we reported the induction of rhesus monkey ESCs along a hepatocyte lineage by a three-step method. The Rhesus monkey ESCs were suspended in DMEM medium supplemented with 10% FBS for 11 days to form embryonic bodies (EB) which contained early endoderm or hepatic cells by expression of early hepatic genes. EB of 11 days were transferred into ECM coated dishes and cultured with aFGF, BMP-4 and OSM at different time points for further studies. After induction by aFGF and BMP-4 for 7 - 10 days, the differentiated cells showed a hepatocyte-like morphology, expressed hepatic marker genes (such as AFP，ALB APOH，G-6-P and TAT) and proteins (AFP，ALB and CK18), and possessed the capacity to storage intracytoplasmic glycogen. After further induction with OSM for 7 - 10 days, some cells shared similar characteristics of matured hepatocytes, such as expression of genes of CYP1B1 and ADH1C, and ICG uptaken, indicating the presence of functional hepatocytes in the differentiated cells.