Differentiation of intestinal epithelial stem cells from parthenogenetic embryonic stem cells

Abstract

Intestinal epithelial stem cells (IESCs) have great potential for the repair of intestinal epithelial injury. However, the amplification of primarily cultured IESCs is relatively difficult. This study aims to obtain IESCs by inducing the differentiation of human parthenogenetic embryonic stem cells (hPESCs). The differentiation of hPESCs into to the definitive endoderm (DE) was performed by using activin A and Wnt3a firstly. Then the differentiation of DE into IESCs was conducted in the presence of epidermal growth factor (EGF). Markers of IESCs, i.e., Musashi 1(Msil) and hairy and enhancer of split 1(Hes1), were monitored dynamically with double immunocytochemical staining and real-time quantitative PCR (qPCR) to identify the differentiation of IESCs. Results showed that Msil+Hesl+ IESCs culminated after a 5-day culture in the presence of EGF and the expression of Msil and Hes1 in cells treated by EGF was 51.3- and 45.38-fold times, respectively, higher than those not treated by EGF. There results demonstrate that IESCs can be generated by inducing the differentiation of hPESCs. Therefore, this study provides a potential source of IESCs for the regeneration of injured intestinal epithelia and the theoretical and experimental basis for the clinical application of hPESCs.