一种应用流式细胞术快速分选活的衰老细胞的方法  

A Rapid Flow Cytometric Method for Sorting of Living Senescent Cells

作　　者：阳晶[1] 黄岚珍[1] 黄希仕 杨飞城[1] 沈珂[1] 蒋晓山[1] 

机构地区：[1]桂林医学院科学实验中心,桂林541004

出　　处：《中国细胞生物学学报》2015年第1期74-78,共5页Chinese Journal of Cell Biology

基　　金：广西医学科学实验中心开放基金(批准号:KFJJ2011-13)资助的课题~~

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1.0μg/m L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β-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(P<0.01)。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。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% of cells sorted from the SEN gate were positive for the senescence-associated β-galactosidase staining, while only 8.01% from the PROL gate （P〈0.01）. Our study provides a rapid method for sorting of living senescent cells based on the flow cytometric technology.

关 键 词：衰老细胞 流式细胞术 细胞分选 衰老相关β-半乳糖苷酶 CNE 2细胞 

分 类 号：Q2-33[生物学—细胞生物学]