The mixed cultures and neuronal-like differentiation of human amnion-derived mesenchymal stem cells

Abstract

Objective To observe growth of basic fibroblast growth factor (bFGF)-induced cultures of human amnion-derived mesenchymal stem cells (HAMs) and differentiation into neuronal-like cells.Comparative observation.Methods Amnia from full-term,uterine-incision delivery were donated by 12 healthy women.HAMs were obtained by cell separation and culture techniques,and were passaged and induced by bFGF.From the third passage,a total of 1 ml HAMs,at a density of 1.0 × 107/L,was separately harvested from six samples,which served as group A.A total of 1 ml HAMs,at a density of 1.0 × 107/L,was harvested separately from the remaining six samples,which served as group B.A total of 0.5 ml from the six samples of group A and 0.5 ml from the six samples of group B were combined to form group C.Differences in cell quantity among the three groups were compared by cell quantification and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) analysis.Expressions of a glial cell marker,neuron-specific enolase,and Nestin were detected in the three groups by immunocytochemistry.Results Cell quantification and MTT analysis of live cells,as well as AMSC absorbance,were significantly greater in group C compared with groups A and B at 18 days of culture (P ＜ 0.05),and no significant difference was observed between groups A and B.Glial fibrillary acidic protein,neuron-specific enolase,and Nestin were expressed in all groups following bFGF induction.Conclusion Mixed AMSC cultures promoted proliferation,and bFGF-induced HAMs differentiated into neuronal-like cells. Key words: Amnion ; Mesenchymal stem cells ; Induction ; Mixed culture