Department of Medicine, University of Alcalá, Madrid, Spain.
Department of Biomedical and Clinical sciences, Division of Children's and Women's Health, Faculty of Health Sciences, Linköping University, Linköping, Sweden.
Methods Mol Biol. 2022;2543:35-44. doi: 10.1007/978-1-0716-2553-8_4.
The frequency of apoptotic cells in a given phenotypically defined population is usually calculated the apoptotic index (AI), i.e., the percentage of apoptotic cells displaying a specific linage antigen (LAg) within a population of cells that remain unfragmented and retain the expression of the LAg. However, this approach has two major limitations. Firstly, apoptotic cells fragment into apoptotic bodies that later disintegrate. Secondly, apoptotic cells frequently lose, partially or even completely, the cell surface expression of the LAg used for the identification of specific cell subsets. The present chapter will describe a flow cytometry method to calculate the apoptotic rate (AR) that takes into account both cell fragmentation and loss of lineage antigen expression on measurement of apoptosis using flow cytometry ratiometric cell enumeration that emerges as a more accurate method of measurement of the occurrence of apoptosis in normal and tumoral cell cultures.
在给定的表型定义群体中，凋亡细胞的频率通常通过计算凋亡指数（AI）来确定，即显示特定谱系抗原（LAg）的凋亡细胞在保持未片段化并保留 LAg 表达的细胞群体中的百分比。然而，这种方法有两个主要的局限性。首先，凋亡细胞会分裂成凋亡小体，然后这些凋亡小体解体。其次，凋亡细胞经常会部分或甚至完全丢失用于鉴定特定细胞亚群的 LAg 的细胞表面表达。本章将描述一种流式细胞术方法来计算凋亡率（AR），该方法考虑了在使用流式细胞术比色细胞计数测量凋亡时细胞片段化和谱系抗原表达丢失的情况，这是一种更准确的测量正常和肿瘤细胞培养物中凋亡发生的方法。
