不同诱导因子对人脐带血非造血干细胞神经分化的影响  

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

作　　者：季凤清[1] 王屹[2] 孙海梅[1] 李荣平[1] 王丹妮[1] 曾晓蓓[1] 赵春礼[2] 王秀琴[1] 杨慧[2] 

机构地区：[1]首都医科大学组织学胚胎学教研室 [2]首都医科大学北京神经科学研究所,北京100069

出　　处：《解剖学报》2007年第2期178-181,共4页Acta Anatomica Sinica

基　　金：北京市优秀人才培养基金资助项目(20042D0501805);北京市教委科技发展计划面上项目(KM200610025005)

摘　　要：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。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-defived nonhematopoietic stem cells was detected by flow cytometry. The HUCB-defived 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-defived 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 factor for neurons and astroglia, and RA ＋ EGF ＋ bFGF are the best for oligodendrocytes.

关 键 词：神经细胞 胶质细胞 流式细胞术 人脐带血 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学]