The Nature of Cellular Damage During Freezing

Cryopreservation damage, or cryoinjury, to any cell is, thus, due to three primary factors: osmotic stress/dehydration, oxidative stress, and intracellular ice formation. During cryopreservation and subsequent thawing, sperm are exposed to a variety of changing osmotic and oxidative conditions to which the cell must respond in order to maintain or recover function. Freezing and thawing sperm result in exposure to osmotic change, extracellular and intracellular ice crystals, production of ROS, and resultant downstream effects on motility, lipid phase change, membrane integrity, mitochondrial function, DNA integrity, cell signaling, metabolism, and apoptotic and necrotic cell death. Many of these negative effects occur mainly during the thawing process owing to the relative stability of the frozen aqueous state. In general, it is not likely that cells surviving low-temperature freezing sustain damage by intracellular ice since this phenomenon is known to cause cell rupture and ensuing lethal effects that are not compatible with cell survival. Both osmotic and oxidative changes have been shown to induce ROS in sperm and contribute to cellular damage that may be lethal or sublethal [19, 21, 37, 44, 45, 49, 50, 53, 54, 56-59],

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