Functional hepatic differentiation from mouse induced pluripotent stem cells in vitro
- VernacularTitle:小鼠诱导型多潜能干细胞定向分化为功能性肝细胞
- Author:
Gang WANG
;
Tong ZHANG
;
Qi ZHANG
;
Guihua CHEN
- Publication Type:Journal Article
- Keywords:
Induced pluripotent stem cells;
Hepatic efficient differentiation
- From:
Chinese Journal of Pathophysiology
1989;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To directly differentiate the induced pluripotent stem cells (IPS cells) into hepatocytes in chemically-defined,monolayer conditions in vitro. METHODS:Based on the knowledge of mammalian hepatic development,we differentiated the IPS cells into hepatic lineage efficiently. The present study involves stepwise transition of IPS cells through a series of media designs to first specify cells towards anterior definitive endoderm,and secondly,to induce hepatic maturation. RESULTS:The IPS cells recapitulated the development of liver in vivo at RNA and protein level. Realtime PCR analysis was performed on cell preparations at various times during the culture process to determine the time course and degree to which the IPS cells differentiated toward a hepatocyte phenotype. Analysis included markers for endoderm-specific gene expression (Sox17; FoxA2,CXCR4),hepatocytespecific gene expression (albumin and AFP),and markers for undifferentiated IPS cells (OCT4 and Nanog). The endodermspecific gene expression increased early while the expression of OCT4 and Nanog decreased and hepatocytespecific gene expression progressively increased over the course of the differentiation program. The resultant hepatocytelike cells(HLCs) showed phenotypic similarities with adult hepatocytes (expressing AFP and albumin) and additionally showed functionality expected to welldifferentiated hepatocytes such as indocyanine green(ICG) uptake and release. The P450 activity of cytochromsome was also positive. CONCLUSION:We successfully construct the IPS cells which efficiently differentiate into hepatocytes and recapitulate the hepatocytic development in vivo. The reproducible and efficient generation of HLCs from IPS cells represents a critical step towards the ultimate goal of producing functional HLCs for clinical use including cell transplantation or bioartificial liver support. Moreover,our system provide a valid way to study the liver development since it is monolayer,serumfree and mimic the in vivo liver development.
