人脐带问充质干细胞体外培养、鉴定及其诱导分化的研究

目的通过体外分离、培养及鉴定人脐带间充质干细胞（humanumbilicalcordmesenchymalstemcells，hUCMSCs），探讨其多向分化潜能。方法取正常足月新生儿脐带，采用组织贴壁培养法分离原代hUCMSCs，观察细胞生长形态。采用流式细胞仪技术检测hUCMSCs细胞表型及细胞周期。通过成神经细胞诱导和成脂肪细胞诱导，鉴定hUCMSCs的多向诱导分化能力。结果成功分离和培养hUCMSCs原代细胞，经流式细胞仪鉴定高表达间质细胞标志CD44和CDl05，阳性率为96．73％和96．10％，整合素受体CD29阳性率为99．53％；低表达造血系标志CD34和CD45阳性率为0．80％和1．91％，人白细胞抗原HLA-DR阳性率为1．41％。细胞周期检测hUCMSCs主要处于G0／G1期。hUCMSCs成神经细胞诱导，出现神经元样细胞；免疫组化检测神经巢蛋白（Nestin）呈阳性表达。hUCMSCs成脂肪细胞诱导，出现空泡样脂肪滴，油红O染色可见脂质沉积。结论hUCMSCs具有多向分化潜能，可跨胚层诱导分化为多种组织类型的细胞。

Culture, Identification and Induction Differentiation of Human Umbilical Cord Mesenchymal Stem Cells in Vitro

Objective To isolate, cultivate and identify human umbilical cord mesenchymal stem cells （hUCM- SCs） in vitro to discuss its multiple differentiation potentials. Methods The umbilical blood of a healthy newborn was i- solated from primary hUCMSCs by tissue adherent culture method, and the form of ceils growth was also observed. The cell phenotype and cycle of hUCMSCs were detected by flow cytometry technique. The multiple induction and differentia- tion ability of hUCMSCs were identified by induced neuroblast and lipoblast. Results Primary cells were isolated and cultured successfully, and positive rates in markers of high expression for mesenchymal cell identified by flow cytometer were CD44 （96.73 % ）, CD105 （96.10 % ） and integrin receptor CD29 （99.53 % ） ; positive rates in hematopoietic lineage markers of low expression were CD34 （0.80 % ） and CD45 （ 1.91% ） , and human leucocyte antigen HLA-DR （1.41%）. HUCMSCs were mainly in G0/Gt period by detection of cell cycle. Neuron-like cells emerged in hUCMSCs after nenroblast induction; immunocytochemistry detection showed a positive expression of Nestin protein. Vacuolar lipid droplet emerged after lipoblast induction, and oil red O staining showed a large quantity of lipid depositions. Conclusion HUCMSCs have multiple differentiation potentials, and can be induced to differentiate into ceils of various histological types with across embryonic layer.