不同诱导因子对人脐带血非造血干细胞神经分化的影响

目的 探讨诱导人脐带血非造血干细胞向神经细胞分化的最佳条件.方法 通过密度梯度离心法原代培养脐带血非造血干细胞;用流式细胞术分析脐带血获得的贴壁细胞的CD34、CD90、CDl66、HLA-ABC、HLA-DR等免疫表型;并用不同诱导因子组合:1.维甲酸(RA);2.表皮生长因子(EGF) 碱性成纤维细胞生长因子(bFGF);3RA EGF bFGF;4二甲基亚砜(DMSO) 丁羟基茴香醚(BHA)诱导后,用免疫荧光化学法检测各组脐带血非造血干细胞中β-微管蛋白-Ⅲ(β-tubulin-Ⅲ)、胶质纤维酸性蛋白(GFAP)、半乳糖苷酶-C(Gal-C)等神经细胞、星型胶质细胞及少突胶质细胞的特异性标记抗原的表达.结果 脐带血非造血干细胞CD90、CD166、HLA-ABC表达阳性;无论那一种诱导因子组合,分化为少突胶质细胞的比率均高于神经元及星型胶质细胞;即各组分化为少突胶质细胞数＞神经元的细胞数＞星型胶质细胞数.对于少突胶质细胞(Ga-C)以RA EGF bFGF组合诱导率最高,达57.02%,其次是DMSO BHA诱导组.而对于神经元及星型胶质细胞的分化各组比较,以RA组最高,其次是DMSO BHA诱导组合.结论 RA对神经元及星型胶质细胞的诱导作用较好,而RA EGF bFGF组合对于少突胶质细胞诱导作用最明显.

THE EFFECTS OF DIFFERENT INDUCERS ON THE DIFFERENTIATION OF NONHEMATOPOIETIC STEM CELLS INTO NERVE CELLS IN HUMAN UMBILICAL CORD BLOOD (HUCB)

Objective To explore the best condition for nonhematopoietic stem cells in human umbilical cord blood(HUCB) to differentiate into nerve cells. Methods The mononuclear cell fraction was obtained by Ficoll gradient separation. Immunophenotyping of HUCB derived nonhematopoietic stem cells was detected by flow cytometry. The HUCB derived nonhematopoietic stem cells were stained by immunohistochemical staining after 4 days of teatment with RA, EGF+bFGF, RA+EGF+bFGF and DMSO+BHA. Results The HUCB derived nonhematopoietic stem cells were CD90,CD166 and HLA-ABC positive, but CD34 negative and expressed the neural markers β-tubulin-Ⅲ(neurons), GFAP(astrocytes), and Gal C(oligodendrocytes)after being treated with different inducers. Interestingly, the cells showed a high commitment to oligodendrocytes(57.02%), the positive cell numbers of Gal-C>β-tubulin-Ⅲ>GFAP.Conclusion RA is the best