一种应用流式细胞术快速分选活的衰老细胞的方法-中国细胞生物学学报.PDF

中国细胞生物学学报 Chinese Journal of Cell Biology 2015, 37(1): 74–78 DOI: 10.11844/cj cb.2015.01.0184 一种应用流式细胞术快速分选活的衰老细胞的方法 阳 晶 黄岚珍 黄希仕 杨飞城 沈 珂 蒋晓山* (桂林医学院科学实验中心, 桂林54 1004) 摘要 衰老细胞是细胞老化研究的重要模型, 目前还缺乏从混合细胞群中分选活的衰老细 胞的有效方法。该研究以1.0 μg/mL顺铂诱导的人鼻咽癌CNE 2细胞衰老为研究模型, 根据细胞大 小和自发荧光(脂褐素积累) 两项物理特征, 采用FACS Aria III流式细胞仪在Purity模式下设门分选 β 活的衰老细胞(SEN)和增殖细胞(PROL), 收集细胞, 再分别进行回测分析和衰老相关 -半乳糖苷酶 (SA-β-gal)染色验证。结果从SEN 门控获得分选纯度为74.7%的活的衰老细胞, 分选出的细胞可重 新贴壁; 其中, 72.1%的细胞呈SA-β-gal染色阳性, 与PROL 门控分选获得的细胞(8.01%)相比较有显 著差异(P0.01) 。该研究提供了一种基于流式细胞仪快速分选活的衰老细胞的方法。 β 关键词 衰老细胞; 流式细胞术; 细胞分选; 衰老相关 -半乳糖苷酶; CNE 2细胞 A Rapid Flow Cytometric Method for Sorting of Living Senescent Cells Yang Jing, Huang Lanzhen, Huang Xishi, Yang Feicheng, Shen Ke, Jiang Xiaoshan* (The Center for Science Research, Guilin Medical University, Guilin 541004, China) Abstract Senescent cells provide an important model for the study of aging. However, there is lack of effective methods to isolate living senescent cells from a mixed population of cells. The purpose of this study was to explore a rapid flow cytometric method for high-throughput sorting of living senescent cells. In a cellular senescence model of cisplatin-induced human nasopharyngeal carcinoma CNE 2 cells senescence, we used a FACS Aria III flow cytometry to sort living senescent cells (SEN) and proliferating cells (PROL) based on two physical characteristics of the cell size and autofluorescence (lipofuscin accumulation). A sorting purity of 74.7% of living senescent cells was obtained from the SEN gate in the Purity model by this method. The sorted cells could adhere to growth and 72.1%