免疫磁珠法分选人脑胶质瘤干细胞的体外培养及鉴定

目的:旨在从人脑胶质瘤组织和细胞株中分离脑胶质瘤干细胞、进行体外培养并对其干细胞特性加以鉴定,以期对后续个体化敏感药物化疗奠定基础。方法:采用以CD133为标志的免疫磁珠法从人脑胶质瘤组织和细胞株中分离脑胶质瘤干细胞并进行体外培养,通过免疫荧光技术检测干细胞标志物CD133、Nestin,干细胞特性加以鉴定。结果:新鲜人脑胶质母细胞培养得到悬浮生长的肿瘤干细胞球,其能在体外自我更新、增殖,形成新的克隆性细胞球,保持连续稳定的传代,能表达神经干细胞表面标志物CD133。在含血清培养基中能够分化为与原肿瘤组织相似的贴壁生长的细胞。结论:胶质母瘤细胞株中存在的一小部分CD133+胶质瘤细胞具有干细胞的属性,并能稳定传代增生。

In vitro Culture and Identification of Human Glioma Stem Cells with Immunomagnetic Beads

BACKGROND&OBJECTIVE: To sort brain glioma stem cells(BGSCs) from glioma cell lines and fresh glioma specimens, culture in vitro and identify its character as a stem cell. METHODS: BGSCs were sorted through Magnetic Activated Cell Sorting(MACS, immunomagnetic beads marking by CD133) and cultured in vitro, and character as a stem cell was identified by stem cell markers (CD133 and Nestin)and differentiated cell markers. RESULTS: Only a small subset of CD133+ glioma cells in glioma cell lines and fresh specimens from various pathologic grade could express stem cell markers CD133 and Nestin, view ultrastructure of a stem cell and be capacity of serial passage in culture. they can generate and form tumors that phenotypically resembl the tumor from the patient. CONCLUSION: The small fraction of CD133+glioma cells in glioma cell lines and fresh specimens from various pathologic grade identified stem cells is BGSCs, so called as tumor stem cells in glioma.