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 in vitro. Conclusion. 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.