人骨髓细胞向汗腺细胞诱导培养过程中基因表达方式的研究.docVIP

人骨髓间充质干细胞向汗腺细胞诱导培养过程中基因表达方式的研究 周岗1，谢晓华1，杨靖1，付小兵2，孙同柱2 1 解放军总医院南楼外一科 北京 100853 2 解放军总医院第一附属医院 北京 100037 【摘要】 目的 通过诱导人骨髓间充质干细胞（hMSCs）向汗腺细胞（hSGCs）分化并检测经诱导后细胞的基因表达变化，探讨hMSCs的可塑性分化机制。方法 分离纯化并体外扩增hMSCs，设立对照组（DM组）和向hSGCs诱导的SG组、EGF10组和EGF20组等四组不同的培养基体系，观测细胞形态学变化和采用流式细胞仪、RT-PCR等方法检测融合细胞的基因和蛋白表达特征。结果 细胞在不同的条件培养液中诱导培养均能存活；诱导培养2d后细胞数量开始扩增，细胞形态也由间充质干细胞的长梭状开始向hSGCs的扁平状转变，在SG组尤其明显；诱导培养1周后细胞数量扩增明显；与汗腺发生形成密切的基因EGF和EGFR出现高丰度的表达，并且表达了仅hSGCs才有表达的CK7蛋白。结论 hMSCs在不同介质诱导培养下可在形态学和分子水平上呈现向hSGCs横向分化的潜能，hMSCs的可塑性现象丰富了皮肤创面修复和汗腺再生理论。 关键词：间充质干细胞；汗腺细胞；基因表达 中图分类号：R34；文献标识码：A；文章编号：110246；DOI： 作者简介，，士，副主任医师，E-mail：dr@ Studies on the genes transcriptional changes in the process of inducing human bone marrow mesenchymal stem cells into sweat glands cells ZHOU Gang,1 XIE Xiao-Hua,1 YANG Jing,1 FU Xiao-Bing,2 SUN Tong-Zhu2 1General Hospital of PLA, Postgraduate Medical College, Beijing 100853, P. R. China. 2The First Affiliated Hospital of PLA General Hospital, Beijing 100048, P. R. China. Abstract: Objective: To investigate underlying mechanisms in the plasticity of hMSCs by analysis of the transcriptional changes of genes that occurred in the process of inducing human bone marrow mesenchymal stem cells (hMSCs) differentiate into sweat glands cells (hSGCs). Methods: HMSCs were isolated, purified and proliferated. Control group (DM) and inducing group were set up in the experiment, the latter were subdivided into three groups (SG, EGF10 and EGF20). The phenotype of hMSCs was detected, the percentage of phenotype conversion was calculated, and characteristics of genes and proteins were detected in induced hMSCs using flow cytometry and RT-PCR methods. Results: HMSCs could survive in varied conditional substratum. The numbers of induced passage cells in the defined medium were proliferated slowly in the earlier days and amplified rapidly after one week. Subsets of the induced hMSCs were wide and flatten generally in the phenotype instead of long and spindle, especially in the SG group. Compared with the co