RESUMO
Objective To study the effect of basic fibroblast growth factor(bFGF) on neuron-like differentiation of superparamagnetic iron oxide nanoparticles (SPIONs)-labeled amniotic membrane-derived mesenchymal stem cell.Methods Cells were cultured from enzymatic-digested amniotic membrane tissue.After that,the following steps were taken:(1) Mesenchymal stem cells derived from amniotic membrane were identified by using cell morphology,MTT method and flow cytometry.(2)SPIONs were used to label amniotic membrane-derived mesenchymal stem.(3)bFGF was imported to induce the neuron-like differentiation of SPIONs-labeled amniotic membrane-derived mesenchymal stem cell.Results (1) Primary cultures of P3,amniotic membrane-derived mesenchymal stem cell were fibroblast-like and expression of surface molecules CD29,CD44,CD90 and CD105 was detected,while expression of CD31,CD34,CD45 and CD106 was negative.(2) SPIONs of no more than 14.0 μg/ml are safe to label amniotic membrane-derived mesenchymal stem cells.Cell activity is more than 80％ and expression of surface molecules CD29,CD44,CD90 and CD105 is positive.(3)RT-PCR and immunocytochemistry analysis showed that 10.0 ng/ml bFGF induced neuron-like differentiation of amniotic membrane-derived mesenchymal stem cell (14 μg/ml SPIONs-labeled).Conclusions Enzymatic digestion and cell adherent culture method can be used to isolate mesenchymal stem cells from amniotic membrane.SPIONs of no more than 14.0 μg/ml are safe to label amniotic membrance-derived mesenchymal stem cells and have no effect on the cell activity.Neuron-like differentiation of amniotic membrane-derived mesenchymal stem cell can be induced with 10.0 ng/ml bFGF.
