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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% 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.
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