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.
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.
Keywords: Activin A; Definitive endoderm; Differentiation; Human parthenogenetic embryonic stem cell; Wnt3a.
© 2020 by the Association of Clinical Scientists, Inc.