不同病理级别脑胶质瘤的肿瘤干细胞体外分离培养和增殖分化

Isolation culture and proliferation and differentiation of brain tumor stem cells from distinct grade gliomas in vitro

目的探讨不同病理级别的人脑胶质瘤的肿瘤干细胞在体外分离培养和增殖分化的情况。方法通过对不同病理级别手术切除的胶质瘤新鲜标本在体外无血清培养基中分离培养和体外增殖分化进行观察研究和分组比较它们的增殖能力。结果在无血清培养基中,Ⅰ级胶质瘤的肿瘤干细胞不能形成克隆球;Ⅱ级胶质瘤的肿瘤干细胞可形成中等克隆球;只有Ⅲ-Ⅳ级胶质瘤的肿瘤干细胞才能形成大型克隆球。在血清培养基中,Ⅰ级胶质瘤干细胞增殖缓慢、分化较晚;Ⅲ-Ⅳ级胶质瘤的肿瘤干细胞增殖迅速、快速进入分化期;Ⅱ级胶质瘤肿瘤干细胞增殖分化介于两者之间。分组比较,发现在不同培养基中和不同病理级别胶质瘤干细胞各组之间在增殖和分化能力上均存在统计学差异(P<0.01)。结论胶质瘤的肿瘤干细胞在生物特性上存在多样性。Ⅲ-Ⅳ级胶质瘤的肿瘤干细胞易于分离培养,增殖分化能力较强;Ⅰ级胶质瘤的肿瘤干细胞增殖分化缓慢,有助于动态观察脑肿瘤干细胞自我更新和增殖分化的过程,是一个较好的观察研究对象。

Objective To explore isolation,proliferation and differentiaton of culturing brain tumor stem cells（BTSCs） from distinct grade gliomas in vitro.Methods During the process of isolation,culture and identification in serum-free culture medium,and the process of proliferation and differentiation in serum culture medium in vitro,BTSCs from distinct grade gliomas were observed and compared.Results These phenomenon were observed that BTSCs from grade Ⅰ gliomas could not be cultured to spheroplast,these from grade Ⅱ gliomas can develop to medium-sized spheroplast and these from grade Ⅲ gliomas or grade Ⅳ gliomas can develop to larger spheroplast in serum-free culture medium.As against that of BTSCs from grade Ⅲ gliomas or grade Ⅳ gliomas,proliferation and differentiation of BTSCs from gradeⅠgliomas is slowly in the early stage.Proliferation and differentiation of BTSCs from grade Ⅱ gliomas is between the above two.Proliferation and differentiation of culturing distinct grade BTSCs were compared with cells number among serum culture groups and serum-free culture groups（P0.01）.Conclusion BTSCs from distinct grade gliomas is biologically multifarious.BTSCs from grade Ⅲ gliomas or grade Ⅳ gliomas is easily obstained.Proliferation and differentiation of these BTSCs is faster than those from the other grade gliomas.Because proliferation and differentiation of BTSCs from grade Ⅰgliomas is slowly in the early stage,it is a better experimental subject in the self-renew,proliferation and differentiation of BTSCs.