人脑多形性胶质母细胞瘤原代培养及生长特性观察

Primary culture and growth observation of human glioblastoma multiforme cells

目的探讨原代培养人脑多形性胶质母细胞瘤细胞的方法,观察培养细胞的生长活性,建立人脑多形性胶质母细胞瘤细胞体外实验模型。方法采用酶消化法原代培养14例人脑多形性胶质母细胞瘤细胞,用倒置相差显微镜和荧光显微镜观察增殖过程中细胞的形态学特点,台盼蓝染色法观察生长活性,描记生长曲线以研究细胞的生长特性。结果 12例原代培养成功,第4代以后肿瘤细胞纯度达98%以上。培养成功的细胞状态稳定,增殖活跃,有明显的对数生长期。结论采用酶消化法可成功原代培养人脑多形性胶质母细胞瘤细胞,酶消化法加划除法可获得高纯度的肿瘤细胞,成功率较高,适宜体外培养细胞。

Objective To investigate the method for primary human glioblastoma multiforme（ GBM） cells culture,and establish an experimental model for studying glioblastoma in vitro. Methods GBM cells isolated from 14 glioblastoma patients were cultured primarily by the enzymatic digestion culture method. The cell morphology was observed with inverted phase contrast microscope and fluorescence microscope,and the growth curve was analyzed by trypan blue staining. Results Twelve prmiary cultures of GBM cells were successfully established. The percentage of tumor cells was over 98% after 4 tranfers. The cultured cells had the proliferative ability and logarithmic growth phase. Conclusion The human GBM cells can be cultured and highly purified GBM cells can be obtained by enzymatic digestion culture method.