人骨髓间充质干细胞L型钙离子通道的研究

L-type calcium channels of human mesenchymal stem cells

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。

Objective：To investigate L-type calcium channels of human mesenehymal stem cells（hMSCs） cultured in vitro, Methods： hMSCs were isolated, cultured by Percoll gradient centrifugation, adherence to plastic flask and monoclonal culture, Immunopbenotypes of hMSCs were detected by immunofluorescence and flow eytometry techniques, Whole cell patch-clamp technique was used to observe the change （If L-type calcium channels of human mesenehymal stem cells（hMSCs） , Results： High homogenous hMSCs had been isolated and cultured in vitro. hMSCs had unique immunopbenotypes 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（ICa-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.