改良法体外诱导骨髓基质干细胞分化为类许旺细胞

Experimental study on inducing bone marrow stromal cells into Schwann-liked cells in vitro by modified method

目的 探讨一种较为有效的诱导方法将骨髓基质干细胞体外定向诱导分化为类许旺细胞。 方法 用Dezawa所述方法 (称为传统诱导法 )加以改良成将所有诱导剂一起联合半量间隔诱导两次的方法 (称为改良诱导法 ) ,诱导后的细胞从形态学、抗S 10 0和抗GFAP细胞免疫化学染色的阳性率、MTT细胞活性测定及流式细胞仪检测细胞DNA含量来比较此两种方法对细胞的综合作用。 结果 改良法诱导的细胞在形态上更具有许旺细胞的长梭形外观 ,抗S 10 0及抗 GFAP阳性率分别由5 8 1%和 60 3 %提高到 79 4%、81 2 % (P <0 0 5 ) ;MTT法细胞活性测定表明改良法对细胞活性影响小(P <0 0 1) ;流式细胞仪DNA含量测定显示S期DNA含量较高 (P <0 0 5 )。 结论 改良法体外诱导骨髓基质干细胞为类许旺细胞具有诱导效果更好 ,对细胞损害小的优越性。

Objective To search for a more efficient method to induce bone marrow stromal cells (BMSCs) into Schwann-liked cells (SLCs) in vitro. Methods On the base of the method which was used by Dezawa (it was looked as the traditional inducing method),some steps were modified,namely,to use all the reagents in half dose in the same time and two times intervally,this method was looked as the modified inducing method.After induced by the two methods,the cells morphologic characteristic,the positive ratio of immunocytochemical dye with anti-S-100 and anti-GFAP,the cells activity measured by MTT method and the DNA percentage in S period measured by flow cytometry were compared with each other respectively to evaluate the methods'effects. Results Compared with the traditional method,the cells induced by the modified method were more similar to the primary Schwann cells in morphology,more positive proportion in immunocytochemic dye with anti-S-100 and anti-GFAP,less damage in the activity and more percentage in S period. Conclusion The modified method had more advantages such as less damage on cells and more efficiency in inducing BMSCs into SLCs.