Study on a Cell Viability Detection Method after Flow Cytometric Sorting Based on Real-time Labeling-free Technique

KeyWord:real time cellular analysis  flow cytometric analysis  flow cytometric sorting  CCK-8  cell viability

Abstract:

      In this paper£¬flow cytometric analysis£¬cell counting kit-8£¨CCK-8£© and real time cellular analysis£¨RTCA£© were used to detect cell viability before and after flow cytometric sorting£¬respectively. The experimental operation and statistical analysis of the three methods were compared comprehensively. The result showed that the apoptosis percentages for cells determined by flow cytometric analysis were £¨3.85 ¡À 0.008£©% in the group after sorting and £¨14.09 ¡À 0.021£©% in the group before sorting. And the apoptosis percentage for cells after sorting was lower than that before sorting£¬with a statistical difference£¨P£© less than 0.05. Further statistical analysis for the proportion of living cells before and after sorting showed that the proportion of living cells after sorting was higher than that before sorting£¬with a significant statistical difference£¨P£© less than 0.001. The CCK-8 assay showed that the group after sorting had higher cell proliferation viability than the group before sorting£¨P < 0.001£©. RTCA assay showed a highly significant statistical difference£¨P < 0.000 1£© in the continuous proliferation activity of the cells after sorting. Compared to the assays of flow cytometric analysis and CCK-8£¬RTCA method has a low request of cell numbers£¬recyclable£¬easy operation£¬labeling-free£¬high detection sensitivity£¬which displays a real-time dynamic cell growth curve suitable for studying the long-term dynamic changes of cell viability after sorting. Furthermore£¬RTCA could be used as a method for the functional studies of cells after basic and clinical sorting.