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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-derived stem cells (NTCSCs) from bone marrow of SD rats. Methods NTCSCs were isolated from bone marrow of DMEM / F12 (1: 1) serum-free neural stem cell-conditioned medium. Immunohistochemistry was used to detect the expression of NTCSCs and their differentiation. RT-PCR was used to detect Transcription of nestin, CXCR4, CD31, βⅢ-Tubulin and GFAP mRNA in bone marrow NTCSCs. RESULTS: The spheres were cultured in suspension. Nestin and CXCR4 were expressed in the spheres. The neurons were labeled with βⅢ-Tubulin and glial cell marker GFAP after differentiation. Nestin, CXCR4, CD31 and βⅢ- Tubulin, GFAP gene mRNA transcription. Conclusion NTCSCs were successfully isolated from bone marrow, which has neurobiological features and can be used as seed cells. It has broad application prospect in the treatment of clinical central nervous system diseases and the repair of trauma.