人脑胶质瘤肿瘤干细胞的培养及鉴定

Isolation , Incubation and Identification of Cancer Stem Cell in Human Gliomas

目的:从人胶质瘤中分离并鉴定肿瘤干细胞。方法:用神经干细胞无血清培养方法从胶质瘤中分离出具有单细胞克隆能力的细胞球,通过核型分析证明其肿瘤特性,用MTT法检测单克隆细胞增殖能力,应用免疫组织化学的方法检测克隆细胞nestin抗原和分化后相关神经细胞特异性抗原的表达。结果:所获得的细胞球具有连续克隆的能力,nestin抗原阳性,并具有肿瘤核型,分化后的细胞表达星形胶质细胞和神经元特异性抗原。结论:成功培养出脑胶质瘤肿瘤干细胞,该细胞具有自我更新、多向分化和很强的增殖能力等干细胞的特征。

Objective: To isolate the gliomal cancer stem cells from gliomas. Methods: Tumor sphere cells were isolated from gliomas with serum-free incubationtechniques which used to isolate normal neural stem cells. The karyotype analysis were conducted to demonstrate that the isolated sphere cells were indeed come from tumors. The proliferative capability of these cells were estimated by MTT method, and Expession of nestin protein and specific proteins of neural cell were detected by inmmunohistochemistry staining. Results: The sphere cells which show tumor karyotype had capacity of proliferating and renewing,and immuno-staining with nestin was strong positive, and specific protein of neurons and astrocytes could detected after they differatiated. Conclusion: the cancer stem cell in gliomas whch bad characteristic of normal neural stem cell such as self-renewing ,multiple differatiation and strong proliferative capability were isolated successfully.