【摘要】 目的研究人脑胶质瘤细胞原代培养的方法,观察培养细胞的生长活性与肿瘤级别之间的关系。方法用消化培养法原代培养24例人脑胶质瘤细胞,酶消化法+振荡贴壁法+机械划除法纯化,增殖细胞核抗原(PCNA)免疫组化染色及甲基噻唑基四唑(MTT)法观察培养细胞的生长活性。结果20例原代培养成功,第4代以后肿瘤细胞纯度达98%以上,PCNA、MTT法显示肿瘤细胞的生长活性与肿瘤级别成正比。结论采用消化培养法可成功培养原代胶质瘤细胞,酶消化法+振荡贴壁法+机械划除法的纯化方法可获得较高的肿瘤细胞纯度;肿瘤细胞的生长活性与肿瘤的恶性程度有关,离体细胞的生长活性可作为判断肿瘤恶性程度的一个指标;检测细胞生长活性MTT法比PCNA染色更具敏感性。更多还原

【Abstract】 Objective To investigate the methods of primary human gliomas cell culture and the relationship between the cultured cells and their histological grades.Methods Twenty-four primary gliomas were cultured with enzymatic digestion and the tumor cells were purified with a new protocol. The process was digesting the cells with trypsin at first, then oscillating the cells while they adhered, and mechanically scraping the impurity cells off if necessary at final. In order to observe the cells’ proliferation, methyl thiagolyl tetragolium (MTT) assay and proliferating cell nuclear antigen (PCNA) immunohistochemical staining were applied. Results Twenty primary gliomas were successfully cultured. The new purification protocol could purify the tumour cells beyond 98%. PCNA and MTT assay demonstrated the proliferation of the cultured cells correlated with their histological grades.Conclusion The method of digestion cultivation can successfully culture primary gliomas. Using the purification method of digestion-oscillation-scrape can obtain a high purity of tumor cells. Proliferation of cultured primary tumor cells can be an indicator of the tumor’s biological malignancy. MTT assay is more sensitive than PCNA immunohistochemical staining for observing cells’ proliferation.更多还原