人胎儿脐带间充干细胞培养及其鉴定

Culture and Identification of Human Umbilical Mesenchymal Stromal Cells

目的建立人脐带间充质干细胞(MSC)培养方法,观察其生长情况并进行抗原特异性鉴定.方法获取足月胎儿人脐带,剪碎成组织块,用组织块贴壁法获取MSC.1周时去除组织块,用0.125%胰蛋白酶消化传1～5代.观察形态学变化.用CD44、CD34抗体染色鉴定培养细胞.结果培养第1周,有少量梭形细胞从组织块内迁出,传代后细胞生长加快,细胞形成仿锤形.CD44抗体染色为阳性,CD34染色为阴性,证明其具有MSC特性.结论通过组织块剪碎贴壁法和组织化学染色鉴定获得了MSC,可用于后续研究.

Objective To establish the culture protocol of human mesenchymal stromal cell in vitro,observe the growth of the cells and indentificate them by detecting the specific antigen.Methods Tissues of umbilical cord from human fetus were harvested and tissue blocks were implanted into wells,then incubated into 37 ℃ 5%C02 incubator.After 7 days,the migrated cells from tissue block were digested by using 0.125% trypsin,then passed gradually from l to 5 passages.The morphology of MSC was observed.Lastly,cultured cells were subjected to CD44 and CD34 immunohistochemical staining.Results A few cells migrated out from tissue blocks during the first week.The cells showed a shuttle shape from 1 to 5 passes,with CD44 positive staining and CD34 negative staining.Conclusion MSCs have been isolated by using tissue block adherence wall method and and confirmed with immunohistochemical staining.