摘要
目的 研究碱性成纤维细胞生长因子（bFGF）作为神经元诱导分化剂干预磁标记羊膜间充质干细胞向神经元样细胞分化的效果.方法 酶消化羊膜组织后贴壁培养法培养细胞.（1）采用观察细胞形态、溴化噻唑蓝四氮唑（MTT）法和流式细胞仪鉴定羊膜来源的间充质干细胞.（2）超顺磁性氧化铁纳米颗粒（SPIONs）标记羊膜间充质干细胞.（3）bFGF诱导磁标记羊膜间充质干细胞向神经元样细胞分化.结果 （1）羊膜组织获取的细胞贴壁培养呈成纤维细胞样,P3细胞流式细胞仪检测高表达膜表面蛋白分子CD29、CD44、CD90、CD105.（2）≤14.0 μg/ml SPIONs标记的羊膜间充质干细胞活性大于80％且细胞膜表面蛋白分子阳性表达CD29、CD44、CD90和CD105.（3）PCR和细胞免疫化学染色：10.0 ng/ml的bFGF诱导磁标记羊膜间充质干细胞（14.0 μg/ml SPIONs标记）可向神经元样细胞分化.结论 羊膜组织的酶消化贴壁培养法可获取间充质干细胞;≤14.0 μg/mlSPIONs是羊膜间充质干细胞标记的安全浓度且对细胞生物学特性无影响;10.0 ng/ml bFGF诱导磁标记羊膜间充质干细胞可向神经元样细胞分化.
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.
出处
《中华放射学杂志》
CAS
CSCD
北大核心
2013年第12期1139-1143,共5页
Chinese Journal of Radiology
