The Real-Time Detection of the Effect of Collection Solution on the Viability of Sorted HEK293T Cells
LI Yanwei*, SONG Xinghui, XING Yueting, WANG Jiajia, GUO Chun, HUANG Yingying
Flow cytometry sorting is a commonly used experimental method for purification of living cells and an effective means to obtain highly active cells. The collection solution provides a certain nutrient environment for the sorted cells, thus ensuring the high activity of the sorted cells. This study used flow cytometer to sort human renal tubular epithelial 293T (HEK293T) cells and received them with three groups of collection solution: group A (BSA in PBS buffer), group B (FBS in PBS buffer) and group C (FBS in DMEM medium). Flow cytometry, CCK-8 (Cell Counting Kit-8), and RTCA (Real Time Cellular Analysis) detection methods were used to detect the activity of the sorted cells, and then statistical analysis was performed. The results showed that the cell viabilities of group A (0.4% BSA in PBS buffer), group B (10% FBS in PBS buffer) and group C (10% FBS in DMEM medium) are higher than those of control in each group. The apoptosis rates separately were (4.12±0.474)%, (4.10±0.660)%, (3.71±0.291)%, with significant statistical differences (P<0.000 1). Comparison of the three cell viability detection
methods showed that the RTCA detection method had the advantages of a minimalist operation process, no labeling, and real-time acquisition of the dynamic proliferation curve of cells, which could be used as the most objective detection method for cell viability after sorting. This study provides a reference for revealing the influence of the collection solution on cell viability, and a real-time detection method for rapid and objective detection of cell viability after sorting.
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