BACKGROUND Cryopreservation has been extensively used in assisted reproductive technology, agriculture and conservation programmes for endangered species. The literature reports largely positive results regarding the survival of frozen-thawed cells and clinical outcomes. Nonetheless, it is unclear whether or not cryopreservation of sperm, oocytes and embryos causes any disruption in their genetic integrity. Drawing on the available published evidence, this review paper describes in detail the physical and biochemical factors of cryopreservation that could potentially affect genomic integrity. METHODS A critical review of the published literature using PubMed with particular emphasis on studies which include assessment of genetic stability after cryopreservation of oocyte, sperm and embryos. The search was performed in 2014 and covered the period from the beginning of electronic records until July 2014. No language restrictions were applied. RESULTS Cryopreservation is associated with extensive damage to cell membranes, and results in alteration of the functional and metabolic status of the cells and mitochondria. Some evidence suggests an increase in DNA single-strand breaks, and degree of DNA condensation or fragmentation in sperm after cryopreservation. The extent of these changes may vary between different individuals and different techniques. The addition of antioxidants to the cryopreservation media and the use of well-controlled cooling regimes could potentially improve such outcomes. Limited numbers of studies on oocytes provide controversial results regarding the effect on DNA fragmentation, sister chromatid exchange (SCE) and aneuploidy. The only study on human embryos suggested that vitrification affects DNA integrity to a much lesser extent than slow freezing. Animal studies show increases in mitochondrial DNA mutations in embryos after cryopreservation. The limited numbers of long-term follow-up studies in humans provide reassurance that derives mostly from retrospective studies with some methodological weaknesses. CONCLUSIONS This review provides an overview of studies performed to date on the effect of cryopreservation on the oocyte, sperm and embryos. Controversy of the reported data has highlighted the gaps in our knowledge not only for clinical studies, but also for basic research in human embryos. New perspectives for future research are proposed.

/pdf/the-effect-of-cryopreservation-on-the-genome-of-gametes-and-pisg9trds7.pdf

The effect of cryopreservation on the genome of gametes and embryos: principles of cryobiology and critical appraisal of the evidence

The ability to fully control sexual maturation and spawning and to produce large numbers of high quality seeds ‘on demand’ (i.e. all year long) is a primary requirement for the successful development of aquaculture. This relies on optimal broodstock management practices based on extensive knowledge of the nutritional and environmental requirements of fish in captivity. However, for many established, emerging and new farmed fish species, such knowledge is limited or not available yet. The level of domestication also plays an essential role as stocks with improved traits in farming conditions are selected. Importantly, reliable indicators of egg quality are still lacking and hatcheries still rely on wild harvested broodstocks in many farmed fish species. These key challenges must be addressed urgently to ensure the sustainable development of the European fish farming sector. This review gives an overview of methods to assess egg/sperm quality and many of the most important factors affecting gamete production and quality, including broodstock nutrition, environmental and spawning induction protocols, genetic management, gamete preservation and new reproductive strategies.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/raq.12025

Gamete quality and broodstock management in temperate fish

http://aquagamete.webs.upv.es/wp-content/uploads/2014/10/Cabrita-et-al.-Aquaculture-2014.pdf

Factors enhancing fish sperm quality and emerging tools for sperm analysis

Cryopreservation has facilitated advancement of biological research by allowing the storage of cells over prolonged periods of time. While cryopreservation at extremely low temperatures would render cells metabolically inactive, cells suffer insults during the freezing and thawing process. Among such insults, the generation of supra-physiological levels of reactive oxygen species (ROS) could impair cellular functions and survival. Antioxidants are potential additives that were reported to partially or completely reverse freeze-thaw stress-associated impairments. This review aims to discuss the potential sources of cryopreservation-induced ROS and the effectiveness of antioxidant administration when used individually or in combination.

The roles of reactive oxygen species and antioxidants in cryopreservation.

Hormonal manipulations for the enhancement of sperm production in cultured fish and evaluation of sperm quality

The influence of certain aminoacids and vitamins on post-thaw fish sperm motility, viability and DNA fragmentation.

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The influence of certain aminoacids and vitamins on post-thaw fish sperm motility, viability and DNA fragmentation.