人异柠檬酸脱氢酶1（IDH1）R132 H突变型脑胶质瘤细胞的原代培养及鉴定

摘要

Objective:To establish a simple,convenient and efficient method of culture for human IDH1 mutant primary glioma cells,to provide cell model for the study of glioma harboring IDH1 R132H.Methods:Tissue samples were taken from patients with low-grade glioma,and using tissue cultivation for primary cell culture.The morphology was inspected by inverted microscope.Hematoxylin-eosin staining (HE )and immunohistochemical staining were used to determine the pathological type of glioma.The type of IDH1 mutation was detected by the base sequence anal-ysis.The growth curve were analyzed to investigate the growth character of IDH1 mutant cells in vitro culture.Re-sults:Successfully established the IDH1 R132H mutant astrocytoma cell line and transferred.Conclusion:Normal hu-man IDH1 R132H mutant astrocytoma cells could be obtained by proper methods.%目的：建立简单、方便、高效的异柠檬酸脱氢酶1（IDH1）突变型胶质瘤原代细胞体外培养方法，为研究IDH1突变型脑胶质瘤提供细胞模型。方法：样本组织取自低级别脑胶质瘤手术患者，采用组织块法行原代细胞培养，倒置显微镜下观察细胞形态特点；瘤组织应用苏木精－伊红染色法（HE ）和免疫组织化学染色确定胶质瘤病理类型以及级别；原代细胞提取基因组行碱基序列测序以鉴定IDH1突变类型；采用MTT法绘制生长曲线，检测IDH1突变型细胞的体外增殖特性。结果：成功建立WHOⅡ级IDH1 R132H突变型人脑星形胶质细胞瘤细胞株，并可传代。结论：应用合适的方法以及培养液，可以培养出IDH1 R132H突变型人脑胶质瘤原代细胞。