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BACKGROUND:Cultures from multiple portions of umbilical cord blood mesenchymal stem cells have been shown to undergo more rapid proliferation and attachment than single portions. OBJECTIVE:To observe growth of basic fibroblast growth factor(bFGF)-induced cultures of human amnion-derived mesenchymal stem cells(AMSCs) and differentiation into neuronal-like cells. DESIGN,TIME AND SETTING:Comparative observation.The study was performed at the Laboratory of Microbiology and Immunology,Basic Medical School of Zhengzhou University from January to May 2008. METHODS:Amnia from full-term,uterine-incision delivery were donated by 12 healthy women. AMSCs were obtained by cell separation and culture techniques,and were passaged and induced by bFGF.From the third passage,a total of 1 mLAMSCs,at a density of 1.0×10~4/mL,was separately harvested from six samples,which served as group A.A total of 1 mLAMSCs,at a density of 1.0×10~4/mL,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. MAIN OUTCOME MEASURES: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.Expression of a glial cell marker,neuron-specific enolase,and nestin was 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 AMSCs differentiated into neuronal-like cells.
BACKGROUND: Cultures from multiple portions of umbilical cord blood mesenchymal stem cells have been shown to undergo more rapid proliferation and attachment than single portions. OBJECTIVE: To observe growth of basic fibroblast growth factor (bFGF) -induced cultures of human amnion-derived mesenchymal stem DESIGN (AMSCs) and differentiation into neuronal-like cells. DESIGN, TIME AND SETTING: Comparative observation. The study was performed at the Laboratory of Microbiology and Immunology, Basic Medical School of Zhengzhou University from January to May 2008. METHODS: Amnia from full -term, uterine-incision delivery were donated by 12 healthy women. AMSCs were obtained by cell separation and culture techniques, and were passaged and induced by bFGF. Getting the third passage, a total of 1 mL AMSCs, at a density of 1.0 × 10 ~ 4 / mL, was separately harvested from six samples, which served as group AA total of 1 mL AMSCs, at a density of 1.0 × 10 ~ 4 / mL, was harvested separately from the remaining six samples, which serv ed as group BA 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. MAIN OUTCOME MEASURES: 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. Expression of a glial cell marker, neuron-specific enolase, and nestin was detected in the three groups by immunocytochemistry. 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 AMSCs differentiated into neuronal-like cells.