人脐带间充质干细胞在不同孔径聚碳酸酯膜上的生长及迁移  

Growth and migration of umbilical cord mesenchymal stem cells on polycarbonate membrane with different pore sizes

作　　者：李东杰[1] 柴家科[1] 韩焱福[1] 孙天骏[1] 邓虎平[1] 赵京玉[1] 刘玲英[1] 

机构地区：[1]解放军总医院第一附属医院烧伤整形科,北京100048

出　　处：《中华医学杂志》2011年第10期699-702,共4页National Medical Journal of China

基　　金：卫生部卫生公益性行业科研专项经费项目（200802066）

摘　　要：目的观察人脐带间充质干细胞（hUCMSCs）在不同孔径Transwell插件底部聚碳酸酯膜上的生长、迁移情况，探讨对hUCMSCs进行非直接接触式共培养诱导分化时选择适宜膜孔径Transwell插件的依据。方法体外分离、培养hUCMSCs，以丝裂霉素C处理后接种于常用的0．4、3．0和8．0μm三种膜孔径的6孔板Transwell插件中，培养7d，观察、计数三种孔径插件底部贴壁细胞，计算迁移率，并在扫描电镜下观察细胞在多孔膜上的生长、迁移情况。结果培养7d后，0．4、3．0和8．0μm三种插件内贴壁hUCMSCs的迁移率分别为0、1．8％、8．0％，0．4μm孔径迁移率与其他两组相比差异有统计学意义（P〈0．01）。扫描电镜下观察到细胞在3．0和8．0μm孔径聚碳酸酯膜底面生长以及穿越微孔的现象，而在0．4μm孔径膜则未发现。结论3．0和8．0μm孔径聚碳酸酯膜允许hUCMSCs迁移穿越，而0．4μm孔径聚碳酸酯膜则不允许，或可利用其正反面进行近距离非直接接触式共培养诱导hUCMSCs分化。Objective To observe the growth and migration of human umbilical cord mesenchymal stem cells （hUCMSCs） on polycarbonate membrane with different pore sizes and explore the criteria of selecting optimal Transwell insert for indirect co-culture to induce the differentiation of hUCMSCs. Methods hUCMSCs were isolated in vitro and then expanded in culture medium. After the treatment of mitomycin C, the cells were seeded on porous membranes of 6-well-dish Transwell inserts with different pore sizes of 0. 4, 3.0 and 8. 0 μm respectively. After culturing for 7 days, the cells were observed and counted on the bottom of each porous membrane. Then the calculation of migration ratio was performed. The growth and migration of hUCMSCs on porous membranes were also examined under scanning electron microscope （SEM）. Results The migration ratios of hUCMSCs on membranes of 0. 4, 3.0 and 8. 0 μm pore sizes were 0, 1.8% and 8.0% respectively. The migration ratio of cells on 0.4 μm pore size membrane was statistically different from that of the other two pore size groups （P 〈 0. 01 ）. Under SEM, a small pm tion of cells were growing on the bottoms of membranes and moving through the pores. But there was no cell movement through 0. 4μm pore size membrane. Conclusions hUCMSCs can migrate through the polycarbonate membranes of 3.0 μm and 8.0 μm pore sizes but not through the 0. 4 μm one. Thus both sides of polycarbonate membrane of 0. 4 μm pore size may be used for close indirect co-culture to induce the differentiation of hUCMSCs.

关 键 词：充质干细胞 人类 脐带 细胞分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]