高接种密度对保留人永生化骨髓基质干细胞体内外分化能力的影响  

Effects of high plating densities on in vitro and in vivo differentiation capability of immortalized human mesenchymal stem cells

作　　者：孙泽林[1] 张亚卓[2] 

机构地区：[1]河北联合大学附属医院神经外科,河北省唐山市063000 [2]北京市神经外科研究所

出　　处：《中华医学杂志》2013年第45期3640-3642,共3页National Medical Journal of China

基　　金：国家自然科学基金（30672156）

摘　　要：目的 观察不同接种密度对端粒酶永生化骨髓基质干细胞（hMSC-TERT）体内外分化潜能的影响.方法 从亲代hMSC-TERT中,分别以1：2和1：20传代,建立不同的接种密度的2个独立的细胞系hMSC-TERT2 和hMSC-TERT20.并对其对各细胞系进行体外向脂肪、骨、神经样细胞诱导分化;用免疫组化的方法测定细胞在SCID鼠的体内多向分化潜能.结果 在体外诱导分化中,hMSC-TERT2形成了脂肪灶和钙化结节,hMSC-TERT20只形成了细胞内脂滴没有形成钙化球.在体内分化中,hMSC-TERT2所形成的组织VIM、GFAP和LCA抗体阳性率（89.17%±5.97%、72.5%±10.11%和76.67%±11.15%）明显高于hMSC-TERT2所形成组织的阳性率（0.33%±0.65%、9.58%±4.29%和22.08%±6.95%）,差异均有统计学意义（均P〈0.01）.结论 高接种密度的培养方式培养似能保持更多的hMSC-TERT细胞体内外分化潜能.Objective To determine whether different cell plating densities could influence the potential differentiation of immortalized human mesenehymal stem cells （hMSC-TERT）. Methods Extensive characterization was performed for two independent cell lines derived from parental hMSC-TERT cell line based on different plating densities during expansion in culture ： 1 ： 2 （ hMSC-TERT2 ） and 1 ： 20 （ hMSC-TERT20 ） . Their adipoeytic, osteogenic, neuronal differentiation potential in vitro and muhidirectional differentiation potential in vivo were analyzed by immunohistoehemistry for pathologic tissue markers. Results hMSC-TERT2 formed cellular adipose loci and mineralized tuberculum after in vitro induction while hMSC-TERT20 showed adipoeytes but no adipose foci or mineralized tuberculum. The positive rate of hMSC-TERT2 formed tissues on VIM, GFAP and LCA （89. 17% ± 5.97% , 72. 5% ±10. 11% and 76. 67%± 11.15% ） under SCID murine skin were more than that of hMSC-TERT20 formed tissues （0. 33% ± 0. 65% , 9. 58%±4. 29% and 22. 08% ±6. 95% ）. Conclusion The cell lines derived at high plating density may have more powerful differentiation potential in vitro and in vivo than those derived at low cell plating density.

关 键 词：端粒酶 基质干细胞 诱导分化 体内分化 接种密度 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]