Role of Wnt5a in the differentiation of human embryonic stem cells into endometrium-like cells.

ABSTRACT

OBJECTIVE: To explore the effects of Wnt5a and Wint7a on the differentiation of human embryonic stem cells (hESCs) into endometrium-like cells, and provide a basis for establishing endometrium-like cell models and a cell source for carrying out further endometrium-related experiments. METHODS: The hESCs established by our center were differentiated into endometrium-like cells in 4 different media including Wnt5a (Group A), Wnt7a (Group B), secreted frizzled related protein (sFRP, an inhibitor of Wnt signal pathway, Group C) and medium alone (Group D). In the differentiated terminal cells, the expressions of cytokeratin (CK) and vimentin were detected with immunofluorescence, and the mRNA levels of CK18, epithelial cell adhesion molecule (EPCAM), estrogen receptor (ER) and progesterone receptor (PR) were determined with RT-PCR. At the same time, the differentiated terminal cells were incubated in medium containing medroxyprogesterone followed by determination of prolactin (PRL). RESULTS: RT-PCR indicated that mRNA levels of CK18, EPCAM, ER and PR were significantly higher in Group A (Wnt5a) than in other groups (all P<0.05), but were significantly lower in Group C (sFRP2) than in other groups (all P<0.05). The changing trend of PRL mRNA was consistent with that of above genes in the 4 groups. Immunofluorescence displayed that the expression of cytokeratin was the strongest in Group A (Wnt5a), and the weakest in Group C (sFRP2) among the 4 groups. CONCLUSION: Wnt5a has promotive effects on the differentiation of hESCs into endometrium-like cells, but Wnt7a has no marked effects.