Myogenic differentiation of human amnotic fluid-derived mesenchymal stem cells induced by 5-Aza-2'-deoxycytidine in vitro

Abstract

Objective To investigate the efficacy of 5-AzaC to induce the myogenic differentiation of human amniotic fluid-derived mesenchymal stem cells (HAFDMSCs) in vitro. Methods HAFDMSCs were isolated from second trimester amniotic fluid and cultured in vitro. Osteogenic and adipogenic differentiation was carried out to study multipotent potential of HAFDMSCs. HAFDMSCs were cultured in myogenic differentiation medium temporarily supplemented by 10 ummol/L 5-AzaC for 24hours or complete medium as experimental and control groups, respectively. The differentiated cells were identified morphologically and by the expressions of myocyte specific proteins after induction for 2 weeks. Results HAFDMSCs grew well in vitro and didn't reach the plateau phase even after being cultured for 7 days. Adipogenic and osteogenic differentiation potential of HAFDMSCs was confirmed by Alizarin red and oil red O staining. After 5-AzaC induction, HAFDMSCs appeared elongated shape.Immunofluorescence staining and RT-PCR confirmed the expression of myocyte specific phenotypes like Desmin,Tn I and α-Actinin. Conclusions 5-AzaC can induce the HAFDMSCs to differentiate into myoblast-like cells in vitro. Key words: Stem cells,mesenchymal; Cell differentiation; Skeletal muscle