动物细胞大规模培养中细胞凋亡检测方法的研究

为了探讨动物细胞大规模培养中细胞凋亡的检测方法,以不同浓度氯化铵诱导BHK-21细胞凋亡,每隔12 h收集细胞,采用台盼兰形态学排染法、吖啶橙和溴化乙锭双重荧光染色法、FITC-Annexin V/PI双染色荧光显微镜观察法、原位缺口末端标记法(TUNEL)、DNA凝胶电泳、流式细胞术(FCM)法进行细胞凋亡的检测。通过比较发现,不同方法检测结果都存在着显著性差异,但吖啶橙和溴化乙锭双重荧光染色法是一种简单、易行、准确、全面定性细胞凋亡的较为可靠的方法,而FITC-Annexin V/PI双染色流式细胞仪能特异地、准确地检出早期凋亡的细胞,是较为理想的凋亡定量检测方法。这2种方法的联合运用更适应于动物细胞大规模培养过程中细胞凋亡的检测。

Different Methods of Detecting Animal Cell Apoptosis in Large-scale Culture

In order to investigate the detection methods of the cell apoptosis in large-scale culture, with different concentrations chloride inducing BHK-21 cell apoptosis, collecting cells every 12 hours. Using the methods of trypan blue morphology, acridine orange and ethidium bromide double fluorescent staining, FITC-Annexin V/PI double cytometry to detect cell apoptosis. The results showed that the method of acridine orange and fluorescence microscope observation, in situ Nick-End labeling, DNA gel electrophoresis, flow cytometry to detect cell apoptosis. The results showed that the method of acridine orange and ethidium bromide double fluorescent staining is a simple and easy and accurate, comprehensive qualitative cell apoptosis relatively reliable method, and FITC-Annexin V/PI double dyeing flow cytometric is the ideal apoptosis quantitative detection methods, to specific, accurate method of detecting cell apoptosis. Therefore, according to the actual situation uniting and selecting the method flexibly in the study of cell apoptosis.