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Apoptosis is not the primary pathway of cell death after a freeze-thaw cycle, but its induction occurs in a significant part of mussel larval cells (up to 24%). Unlike sea urchin cells , we found that 5% dymethyl sulfoxide without any additives provided the maximum level of cell survival according to the MTT assay, trypan blue exclusion test, and DAPI staining to estimate cell viability.
When the cells are frozen, the glycerol freezes very uniformly without much formation of water ice crystals that could swell and burst the cell. In addition, the cells are frozen at very low temperatures (minus 122.80F or lower). This places the cells in a state of suspended animation.