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Advances in the detection methods for assessing the viability of cryopreserved samples
doi: 10.1515/fzm-2025-0012
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Abstract: Since the beginning of the 21st century, modern medical technology has advanced rapidly, and the cryomedicine has also seen significant progress. Notable developments include the application of cryomedicine in assisted reproduction and the cryopreservation of sperm, eggs and embryos, as well as the preservation of skin, fingers, and other isolated tissues. However, cryopreservation of large and complex tissues or organs remains highly challenging. In addition to the damage caused by the freezing and rewarming processes and the inherent complexity of tissues and organs, there is an urgent need to address issues related to damage detection and the investigation of injury mechanisms. It provides a retrospective analysis of existing methods for assessing tissue and organ viability. Although current techniques can detect damage to some extent, they tend to be relatively simple, time-consuming, and limited in their ability to provide timely and comprehensive assessments of viability. By summarizing and evaluating these approaches, our study aims to contribute to the improvement of viability detection methods and to promote further development in this critical area.
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Key words:
- cryomedicine /
- rewarming /
- tissues and organs /
- viability /
- detection
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Table 1. The differences between various apoptosis detection assay methods
Detection method Detection object Time-consuming Applicable scenarios Annexin V/PI Early apoptotic cells and late apoptotic cells 2-4 h Flow cytometry-based apoptosis staging; drug screening & apoptosis mechanism studies TUNEL Apoptotic cells 6-24 h Histopathological analysis; in vivo/in vitro apoptosis detection Detection of caspase-3 viability Early apoptotic cells 1-2 h Early apoptosis event studies; apoptotic signaling pathway validation Lactate dehydrogenase (LDH) release assay Cell membrane damage 2-4 h High-throughput cytotoxicity testing 
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Table 2. The differences between various cell viability assays methods
Detection method Detection object Time-consuming Applicable scenarios MTT Metabolic activity of living cells 4-6 h High-throughput drug screening; cell proliferation & cytotoxicity studies Neutral red uptake assay Lysosomal activity & membrane integrity of living cells 3-4 h Cytotoxicity assessment (chemical/environmental toxins); long-term culture viability monitoring Western blot assay for β-actin protein Cytoskeletal protein expression (internal reference) 12-24 h Cell structural integrity evaluation; experimental normalization control Calcein-AM cell viability assay Esterase activity in viable cells 30 min-1 h Real-time live-cell imaging; rapid viability assessment MTT, 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide.
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