Efficient Definitive Endoderm Differentiation from Human Parthenogenetic Embryonic Stem Cells Induced by Activin A and Wnt3a.

Objective: This study aimed to investigate the effects of combined activin A and Wnt3a treatment on definitive endoderm (DE) differentiation from human parthenogenetic embryonic stem cells (hPESCs).

Methods: hPESCs on human foreskin fibroblast feeder layers were induced to differentiate into DE using a combination of 50 ng/ml activin A and 25 ng/ml Wnt3a. Expression of the DE markers CXCR4, E-cadherin (ECD), Sox17, and Goosecoid (Gsc) were examined using flow cytometry and real-time quantitative PCR.

Results: The combination of activin A and Wnt3a significantly enhanced the percentages of CXCR4, ECD, Sox17, and Gsc cells, culminating on day 2 of induction. This combined use promoted DE differentiation from hPESCs .

Conclusions: Through the combination treatment using activin A and Wnt3a, DE differentiation from hPESCs culminated at 48 h, which can be regarded as the optimal time-point to induce differentiation of endodermal cells such as pancreatic, liver, and intestinal cells.