2022, Vol. 26 ›› Issue (24): 3870-3874

In vitro differentiation of human placenta-derived mesenchymal stem cells into hepatocytes

Liang Tingting1, Chen Li2, Fan Mingsong2, Zhang Guoying2, Zhang Shichang1   

Abstract: BACKGROUND: Mesenchymal stem cells are an attractive source of therapies for liver failure. The differentiation of mesenchymal stem cells into hepatocytes can improve the therapeutic effect of liver failure.  
OBJECTIVE: To investigate the feasibility of placenta derived mesenchymal stem cells to differentiate into mature hepatocytes in vitro for the treatment of liver failure.
METHODS: Placenta-derived mesenchymal stem cells were isolated and cultured from human placenta tissue. The surface markers and differentiation potential of the isolated cells were identified by flow cytometry, and multidirectional induction. Placenta-derived mesenchymal stem cells were induced into hepatocytes by several cytokines for 22 days. Hepatic markers of induced cells were detected by immunofluorescence staining. Glycogen staining was used to detect the glycogen synthesis ability of cells.  
RESULTS AND CONCLUSION: (1) Under the inverted microscopy, the cultured cells were spindle or fibroblast-like cells, which proliferated rapidly. There was no obvious change in cell morphology after 10 passages. (2) Flow cytometry results revealed that the positive rate of CD73, CD90, CD105, CD44, CD166, CD29, CD54, and HLA-ABC in placenta-derived mesenchymal stem cells was more than 95%, while CD34, CD45, CD117, CD14, CD19, CD133, CD31, and HLA-DR were all negatively expressed. (3) The results of oil red O, alizarin red, and alcian blue staining were positive after adipogenic, osteogenic and chondrogenic differentiation of placenta-derived mesenchymal stem cells. (4) After 22 days of hepatic differentiation, these cells exhibited typical morphology of hepatocytes such as round or polygonal shape with obvious cell boundary. Immunofluorescence staining showed that a large number of differentiated cells expressed albumin, CK-18, HepPar1, and CYP7A1 after induction. The positive staining of differentiated cells was observed by glycogen staining. (5) Our results demonstrated that the cells isolated from placenta were mesenchymal stem cells, and placenta-derived mesenchymal stem cells can differentiate into mature hepatocytes in vitro, which could provide reliable sources of hepatocytes for the treatment of liver failure.
Key words: stem cells, mesenchymal stem cells, placenta, hepatocytes, liver failure, mature hepatocytes, in vitro induction