人早期胚胎间充质干细胞在新生小鼠肾脏内的存活和分布  

Distribution and survival of human embryonic mesenchymal stem cells in kidney of newborn mice

作　　者：袁莉[1] 仵敏娟[2] 张懿[1] 孙红玉[2] 熊俊[2] 刘春艳[1] 刘厚奇[2] 梅长林[1] 

机构地区：[1]第二军医大学长征医院肾内科解放军肾脏病研究所,上海200003 [2]第二军医大学组胚教研室

出　　处：《中华肾脏病杂志》2009年第9期683-687,共5页Chinese Journal of Nephrology

基　　金：基金项目：上海市重点学科建设项目（B902）

摘　　要：目的从早期人胚分离和培养人早期胚胎间充质干细胞（hMSC），鉴定其生物学特性以及分化能力，观察hMSC在新生小鼠肾脏内存活和分布情况。方法取4～7周胚龄的人胚，分离组织进行培养，免疫组化和流式细胞术检测hMSC的生物学特性，以及成骨、成脂肪的分化能力。激光共聚焦显微镜观察PKH-26标记的hMSC在新生鼠肾脏的存活及分布情况。结果分离培养得到的hMSC呈纤维状，贴壁生长，增殖能力强。流式细胞仪检测显示细胞不表达CD34、CD45，但明显表达CD29、CD44、CD90。细胞免疫组化结果显示细胞表达SH-2、OCT-4。在成脂肪和成骨诱导培养的条件下具有多向分化的能力。观察1个月，PKH-26标记的hMSC在新生鼠肾脏中仍存活，10．0％±2．1％分布在。肾小管组织。结论从早期人胚得到的hMSC有较强的增殖和分化能力，免疫力低，在小鼠肾脏内能长期存活，并可能参与小管上皮细胞的发育。Objective To determine the biological characteristics of human embryonic MSC （hMSCs） and their potential ability of differentiation, and to investigate the survival and distribution of hMSCs after transplantation into the kidney of newborn mice. Methods hMSCs were derived from 4-7 week-old embryos, then primary culture was done. The biological characteristics of hMSCs were detected by immunohistochemical methods and flow cytometry. Their differentiation potential was determined by cocuhure with conditioning medium. The survival and distribution of PKH-26-stained hMSCs in mice were observed by laser scanning eonfoeal microscope. Results Flow eytometry and immunochemistry staining revealed that the expression of CD29, CD44, CD90, SH-2, OCT-4 was positive significantly, and CD34, CD45 was negative. The cells could be induced to differentiate to osteocytes and adipocytes under special conditions. After transplantation for 1 month, PKH-26-stained hMSCs still existed in the kidney of mice and co-localized in tubular epithelium by confoeal microscope. Conclusion hMSCs derived from the early human embryo have the ability of proliferation and differentiation with low immunity, and may be involved in the development of renal tubule in newborn mice.

关 键 词：间质干细胞 胚胎 肾小管 上皮细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]