骨髓源性神经组织干细胞的分离、培养及鉴定  

Isolation,culture and identification of neural tissue committed stem cells from bone marrow

作　　者：李琳[1] 张兴秀[2] 王健[2] 姜容[1] 郑敏[1] 

机构地区：[1]重庆医科大学干细胞与组织工程研究室组织学与胚胎学教研室,重庆400016 [2]重庆医科大学附属第二医院神经内科,重庆400016

出　　处：《中国生物制品学杂志》2012年第4期506-509,共4页Chinese Journal of Biologicals

基　　金：国家自然科学基金(31100985);重庆市科委自然科学基金(CSTC;2010BB5096)

摘　　要：目的分离、培养并鉴定SD大鼠骨髓中神经组织干细胞(Neural tissue committed stem cells,NTCSCs)。方法采用DMEM/F12(1∶1)无血清神经干细胞条件培养液,从大鼠骨髓中分离、培养克隆生长的NTCSCs,采用免疫组化法对骨髓NTCSCs及其分化进行鉴定,RT-PCR法检测骨髓NTCSCs中Nestin、CXCR4、CD31、βⅢ-Tubulin、GFAP基因mRNA的转录。结果分离培养获得悬浮生长的细胞球,该细胞球表达神经干细胞标志Nestin及CXCR4,分化后表达神经元标志βⅢ-Tubulin和神经胶质细胞标志GFAP;该细胞中有Nestin、CXCR4、CD31、βⅢ-Tubulin、GFAP基因mRNA转录。结论在骨髓中成功分离到NTCSCs,其具有神经生物学特征,可作为种子细胞,在临床中枢神经系统疾病的治疗及创伤的修复中具有广阔的应用前景。Objective To isolate, culture and identify neural tissue committed stem cells （NTCSCs） from bone marrow of SD rats. Methods NTCSCs were isolated and cultured from rat bone marrow by using serum-free DMEM/F12 （1 ： 1 ） conditional medium, then identified for differentiation by immunohistochemical assay, and determined for transcriptions of Nestin, CXCR4, CD31, [3 m- Tubulin and GFAP mRNAs by RT-PCR. Results Cell spheres in suspension were isolated and cultured, in which Nestin and CX- CR4 as markers of neural stem cells were expressed. β Ⅲ-Tubulin as neuron marker and GFAP as gliacyte marker were expressed in the cells after differentiation. The transcriptions of Nestin~ CXCR4, CD31, β Ⅲ-Tubulin and GFAP mRNAs were proved in the cells. Conclusion NTCSCs with neurobiological characters were successfully isolated from bone marrow and might be used as cell seeds, which provided a basis for therapy of diseases and repair of wound in central nervous system.

关 键 词：骨髓 神经组织干细胞 细胞增殖 细胞分化 

分 类 号：Q254[生物学—细胞生物学] Q78