人脐血间充质干细胞体外诱导分化为类雪旺细胞的初步研究

Experimental Study on the Differentiation of Human Umbilical Cord Blood Mesenchymal Stem Cells into Schwann Cell-like in Vitro

目的：体外定向诱导人脐血间充质干细胞（HUCBMSCs）,分化为类雪旺细胞（SC-like）.方法：（1）采用Ficoll密度梯度离心法分离健康产妇脐带血中单个核细胞进行体外培养,用流式细胞术检测细胞表达的表面抗原CD90,SH2,CD34和CD45.（2）第3次传代所得的HUCBMSCs,加入加有β-巯基乙醇（β-ME）、全反式黄酸（RA）、Forskolin、b-FGF、PDGF、HRG的含10%胎牛血清（FBS）的低糖DMEM培养基（L-DMEM）诱导,7 d后免疫组织化学染色法检测.结果：（1）HUCBMSCs在体外培养以梭形细胞为主 流式细胞仪检测显示,细胞高表达表面抗原CD90和SH2,低表达表面抗原CD34和CD45.（2）诱导7 d后,细胞免疫组化显示,GFAP阳性率为81.88%±2.43%.结论：在一定条件下,HUCBMSCs可以在体诱导分化为SC-like,组成人工神经,移植修复周围神经缺损.

Objective：To induce the human umbilical cord blood mesenchymal stem cells （HUCBM- SCs） to differentiate into schwann cell-like （SC-like） in vitro. Methods： （ 1 ） Mononuclear ceils were separated from umbilical cord blood of healthy parturients by Ficoll density gradient centrifugation and cultured in vitro. The expression of surface antigens CDg0, SH2, CD34 and CD45 were detected by flow cytometry （FCM）. （2） The third passage cells were cultured in low carbohydrates-Dulbecco ＇s modified eagle ＇s medium （L-DMEM） containing 10% fetal bovine serum（FBS） , β-mercaptoethanol （β-ME） , retinoic acid（RA） , forskolin, basic fibroblast growth factor （b-FGF）, platelet-derived growth factor（PDGF） and heregulin （HRG）. On 7th day, the ceils were identified by immunocytochemistry. Results： （ 1 ） The majority of HUCBMSCs cultured in vitro displayed a spindle shaped appearance. FCM showed that the surface antigens CD90 and SH2 were highly ex- pressed in these ceils, while the CD34 and CD45 were very low. （2） On 7th day, the results of immunocyto- chemistry showed that the cells were positive for GFAP. The positive percentageswere 81.88% -＋ 2.43% re- spectively. Conclusions：The HUCBMSCs can be induced to differentiate into SC-like in vitro,which can consti- tute artificial neural to be transplanted to repair the nervus peripheralis defect.