Programmed cell death detection methods: a systematic review and a categorical comparison

Abstract

Programmed cell death is considered a key player in a variety of cellular processes that helps to regulate tissue growth, embryogenesis, cell turnover, immune response, and other biological processes. Among different types of cell death, apoptosis has been studied widely, especially in the field of cancer research to understand and analyse cellular mechanisms, and signaling pathways that control cell cycle arrest. Hallmarks of different types of cell death have been identified by following the patterns and events through microscopy. Identified biomarkers have also supported drug development to induce cell death in cancerous cells. There are various serological and microscopic techniques with advantages and limitations, that are available and are being utilized to detect and study the mechanism of cell death. The complexity of the mechanism and difficulties in distinguishing among different types of programmed cell death make it challenging to carry out the interventions and delay its progression. In this review, mechanisms of different forms of programmed cell death along with their conventional and unconventional methods of detection of have been critically reviewed systematically and categorized on the basis of morphological hallmarks and biomarkers to understand the principle, mechanism, application, advantages and disadvantages of each method. Furthermore, a very comprehensive comparative analysis has been drawn to highlight the most efficient and effective methods of detection of programmed cell death, helping researchers to make a reliable and prudent selection among the available methods of cell death assay. Conclusively, how programmed cell death detection methods can be improved and can provide information about distinctive stages of cell death detection have been discussed.

Integrating Synthesis, Physicochemical Characterization, and In Silico Studies of Cordycepin-Loaded Bovine Serum Albumin Nanoparticles

“…Compared to MTT assay, factors such as cellular adaptation, treatment degradation/metabolism, and cell proliferation are less likely to directly affect the AO/EtBr dual staining method. The AO/EtBr staining specifically targets the structural and functional changes in cells undergoing apoptosis, regardless of the underlying mechanisms or the specific cellular responses to the treatments . The AO dye stains the nucleic acids of live cells, emitting green fluorescence, while the EtBr dye enters cells with compromised cytoplasmic membranes, such as dead or apoptotic cells, emitting red fluorescence .…”

Human umbilical cord mesenchymal stem cell‐derived exosome suppresses programmed cell death in traumatic brain injury via PINK1/Parkin‐mediated mitophagy

“…Pyroptosis is a type of inflammatory PCD that is characterized by the formation of pores in cell membrane, cell lysis, and release of pro‐inflammatory cytokines 27 . Pyroptosis is primarily induced by inflammasomes and performed by the gasdermin (GSDM) protein family such as GSDMD and caspases such as caspase‐1 29 . Inflammasomes are multimolecular complexes containing pattern‐recognition receptors (PRR), apoptosis‐associated speck‐like protein containing CARD (ASC) and NLRP3 30 .…”

Protective effect of sex steroid hormones on morphological and cellular outcomes after neonatal hypoxia-ischemia: A meta-analysis of preclinical studies

“…Another interesting outcome to be evaluated is activation of cell death pathways. Apoptosis is one of the main mechanisms of cell death following neonatal HI and cysteine-aspartic proteases (caspases), Bax and DNA fragmentation staining techniques are important biomarkers of damage [75]. Several studies suggest that steroid hormones can modulate apoptotic pathways, in ammatory responses, mitochondrial dysfunction, blood-brain barrier disruption and neurogenesis [11,12,27,62,73,74].…”