Department of Immunology, Faculty of Health Science, University of Pretoria, Pretoria, South Africa.
Department of Molecular Medicine and Haematology, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa.
PLoS One. 2022 Jun 29;17(6):e0270599. doi: 10.1371/journal.pone.0270599. eCollection 2022.
Cell death is important in physiology, and can happen as a result of structural damage, or as a sequence of programmed cellular processes known as apoptosis. Pathogenic alterations in apoptosis occur in a number of diseases, including cancer, viral infections, autoimmune diseases, immunodeficiencies, and degenerative conditions. Developing accurate and reproducible laboratory methods for inducing and detecting apoptosis is vital for research into these conditions. A number of methods are employed to detect cell death, including DNA fragmentation, the TUNEL assay, and electron microscopy although each has its limitations. Flow cytometry allows for the distinction between live, early apoptotic, late apoptotic and necrotic cells. In this protocol we successfully induce apoptosis using chemical treatment and treatment with low pH in solid tumour cell lines, and have optimized detection using the Annexin V/PI apoptosis assay.
细胞死亡在生理学中很重要，它可能是由于结构损伤引起的，也可能是一系列称为细胞凋亡的程序性细胞过程的结果。细胞凋亡的病理性改变发生在许多疾病中，包括癌症、病毒感染、自身免疫性疾病、免疫缺陷和退行性疾病。开发用于诱导和检测细胞凋亡的准确且可重复的实验室方法对于这些疾病的研究至关重要。许多方法可用于检测细胞死亡，包括 DNA 片段化、TUNEL 检测和电子显微镜检查，但每种方法都有其局限性。流式细胞术可区分活细胞、早期凋亡细胞、晚期凋亡细胞和坏死细胞。在本方案中，我们成功地使用化学处理和低 pH 值处理诱导固体肿瘤细胞系中的细胞凋亡，并通过 Annexin V/PI 凋亡检测优化了检测。
