脐带组织间充质干细胞分离及向成骨与脂肪细胞的分化

目的探讨人脐带组织间充质干细胞体外分离、扩增与成骨、成脂肪细胞诱导分化的方法与条件。方法取脐带沿血管长轴切开,去掉血管,再将脐带重新缝合形成环状,灌入胶原酶悬液,6～8h后灌洗离心,获取细胞培养、扩增,细胞呈集落生长后传代,取传代细胞进一步行免疫表型测定和成骨、成脂肪细胞诱导分化。结果去除血管后获取脐带组织细胞的方法,可获得贴壁生长的细胞,呈短棒状或梭形样细胞,易扩增和形成集落,有基质细胞免疫表型表达,成骨诱导分化的细胞茜素红染色胞浆中有大量的钙沉积,碱性磷酸酶钙钴法染色胞质呈灰黑色,阳性细胞百分率>85%;成脂肪分化的细胞油红染色示胞浆充满了油滴空泡。结论脐带组织存在具有间叶细胞分化能力的间充质干细胞,并可在体外进行培养扩增形成集落细胞传代,传代细胞表达基质细胞表面抗原,能够向成骨细胞、成脂肪细胞分化。

Isolation of Multipotent Mesenchymal Stem Cells from the Tissue of Umbilical Cord for Osteoblasts and Adipocytes Differentiation

Objective To explore the methods and conditions for isolating and proliferating multipotent mesenchymal stem cells(MSCs)from the tissue of umbilical cord,with an aim to induce osteogenic and adipogenic differentiation in vitro.Method The cord was dissected along the long axis,with vessels pulled away and then sutured into a "loop".Collagenase solution was filled into the loop.Suspended cells were collected from the loop suspension after 6-8 hours and centrifuged.The cells were finally cultured in polystyrene dishes.The single cell-derived colonies were obtained and tested for their immunophenotype and osteoblast and lipoblast differentiations.Result Adherent cells were obtained from the tissue of umbilical cord,which proliferated and formed single cell-derived colonies.The colonies presented matrix cells immunophenotype and differentiated into osteoblasts that produced mineralized matrices,which were stained by alizarin red and alkaline phosphatase.The colonies also differentiated into adipocytes that accumulated lipid vacuoles,which were demonstrated by the morphology and oil red stains.Conclusion MSCs can be isolate from the tissue of umbilical cords and proliferate in vitro.The proliferated colonies show matrix cell immunophenotypes and can differentiate into osteoblasts and adipocytes.