期刊
JOURNAL OF PHARMACOLOGICAL AND TOXICOLOGICAL METHODS
卷 124, 期 -, 页码 -出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.vascn.2023.107474
关键词
A549; Cell line; Cell therapy product; Cell viability; Cryoprotection; Freeze-thaw; hMSCs; Methods
The determination of cellular viability is crucial for the safety and effectiveness of frozen cell therapy products. This study evaluated commonly used viability assays in human mesenchymal/stromal stem cells and lung carcinoma cells, and found significant differences in cell viability measurements depending on the assay used.
For the safety and efficacy of frozen cell therapy products, determination of cellular viability is key. However, results of cell viability measurements do not only depend on the cell line or on the inflicted stress, but also on the assay used, making inter-experimental comparisons difficult. The aim of this study was thus to assess commonly used viability assays in clinically relevant human mesenchymal/stromal stem cells and human A549 lung carcinoma cells. Post freeze-thaw stress viability and proliferation were evaluated under different conditions using trypan blue, acridine orange/DAPI stain, alamarBlue, ATP, and neutral red assays. Significant differences in cell viability between metabolic assays were observed, likely due to their distinct intrinsic detection mechanisms. Membrane-integrity based assays generally overestimated cell viabilities in this study. Furthermore, noticeable differences in inter-assay sensitivities were observed. These differences highlight that cell viability methods should be meticulously selected and their associated results carefully interpreted in a relevant context to ensure reliable conclusions. Indeed, although cell membrane integrity based assays are a popular choice to determine cellular quality attributes after freezing and thawing, we demonstrate that metabolic assays may be more suitable in this context.
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