【摘要】 目的:研究分离培养的人骨髓间充质干细胞(hMSCs)膜表面L型钙离子通道的表达。方法:采用密度梯度离心、贴壁筛选及单克隆培养法获得hMSCs,利用流式细胞分析、免疫荧光法鉴定,应用全细胞膜片钳技术记录hMSCs膜表面L钙离子通道的变化。结果:体外分离、培养出高度同源性的具有独特细胞免疫学表型hMSCs;40%的hMSCs表达钙离子电流,峰值电流(ICaPeak)在+20～+30mV为(102.67±19.06)pA,此电流可被Cd2+(20μmolL)阻断。结论:在未分化的hMSCs上L型钙离子通道表达较少,说明L型钙离子通道表达的增多可能是hMSCs定向分化的条件之一,调节hMSCs膜表面L型钙离子通道的表达将有助于促进其定向分化。更多还原

【Abstract】 Objective:To investigate L-type calcium channels of human mesenchymal stem cells(hMSCs) cultured in vitro.Methods:hMSCs were isolated,cultured by Percoll gradient centrifugation,adherence to plastic flask and monoclonal culture.Immunophenotypes of hMSCs were detected by immunofluorescence and flow cytometry techniques.Whole cell patch-clamp technique was used to observe the change of L-type calcium channels of human mesenchymal stem cells(hMSCs).Results:High homogenous hMSCs had been isolated and cultured in vitro.hMSCs had unique immunophenotypes and they were positive for CD44,CD29,c-kit but negative for CD34,CD31,CD54.Calcium currents could be found in 40 percent of hMSCs,peak currents of calcium(I Ca-Peak ) were (102.67±19.06)pA from +20 mV to +30 mV.The currents could be inhibited by Cd~ 2+ (20 μmol/L).Conclusion:Undifferentiated hMSCs express less L-type calcium channes,regulation of the chanels might contribute to directional differentiation.更多还原