汗腺细胞与骨髓间充质干细胞共培养过程中基因表达变化及意义  

Genes expression profile in the co-culture of human bone marrow mesenchymal stem cells with sweat glands cells and the implication

作　　者：周岗[1] 谢晓华[1] 付小兵[2] 杨靖[1] 孙同柱[2] 

机构地区：[1]解放军总医院南楼综合科,北京100853 [2]解放军总医院第一附属医院创伤实验室

出　　处：《中华实验外科杂志》2007年第10期1186-1188,共3页Chinese Journal of Experimental Surgery

基　　金：国家重大基础研究规划资助项目(G1999054204);国家自然科学基金(30500194;30400172)

摘　　要：目的建立人骨髓间充质干细胞(hMSCs)与人汗腺细胞(hSGCs)共同培养模式,检测融合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hSGCs和hMSCs;采用在47℃环境下热休克hSGCs 40min建立hMSCs与hSGCs共培养模式并观测细胞形态学变化;免疫细胞化学双染和RT-PCR等方法检测融合细胞的基因和蛋白表达特征。结果共培养后镜下可见产生融合细胞,细胞形态发生渐变,表现为细胞体积增大,形状不规则,多核和异型核,胞核相互靠近,约占细胞总数的约10%左右。采用抗癌胚抗原(CEA)和抗BrdU免疫细胞化学双染法,检测BrdU标记的hMSCs与hSGCs共培养后的融合细胞,证实该类细胞具有两种细胞的独特的免疫特性;与汗腺发生形成密切的基因EGF和EGFR出现高丰度的表达(P<0.05)。结论热休克共培养后hMSCs具有在形态学和分子水平上向hSGCs分化的潜能,细胞融合现象丰富了皮肤创面修复和汗腺再生理论。To establish the co-culture mode of human bone marrow mesenchymal stem cells （hMSCs） and human sweat glands cells （hSGSs）, study the gene expression profile of the fusion cells and explore the underlying mechanisms of hMSCs differentiation. Methods HMSCs and hSGCs were isolated, purified and expanded respectively. （1-2） x 105 BrdU labeled hMSCs were co-cultured with hSGCs, and the latter were preprocessed with heat shock at 47℃ for 40 min when they reached 70% confluence and then cooled for 1-2 h at 37℃. The phenotype of fusion cells was detected, and characteristics of genes and proteins such as EGF and CEA were tested in the fusion cells by using immunocytochemistry and RT-PCR methods. Results The cultured hMSCs and hSGCs were of clonogenic growth in sweat glands medium. Statistical results showed about 10% of multi-nucleated fusing cells could be found after co-culture for one to two weeks. The phenotype of the fusion cells were wide and flatten. Compared with hMSCs and hSGCs, the expression of genes associated with morphogenesis of sweat glands such as EGF and its receptor EGFR appeared or was enhanced in fusion cells. Some multi-nucleate fusion cells were positive for both anti-CEA and anti-BrdU staining by double-staining immunocytochemistry. Conclusion HMSCs could differentiate into hSGCs at phenotype and molecule level by co-culture of hMSCs with heat shocked hSGCs in vitro. The phenomenon of cell fusing and nuclear reprogramming and hMSCs potential enriched the theory in regulating repair and regeneration of skin and sweat glands.

关 键 词：间充质干细胞 汗腺细胞 培养 基因表达 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]