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Attention to detail and robust quality control systems are essential components for a successful IVF laboratory, necessary to maintain high success rates. This not only pertains to daily laboratory operations, such as adequate staff training, monitoring of equipment function or weekly/monthly success rates, but also entails proper laboratory design and construction. A highly successful IVF laboratory requires careful consideration of layout and workflow and use of appropriate materials to avoid introducing potentially embryotoxic factors into the laboratory environment.
The IVF culture system is constantly being examined for means of modification to further improve conditions for gametes and embryos. Exhaustive research into physiological requirements and responses of these biological cells has provided valuable insight for refinement of culture variables. Extensive testing of conditions, both chemical and physical, has permitted tailoring of the IVF laboratory to the unique requirements of the reproductive cells as well as the needs and preferences of the IVF lab. Furthermore, as with many fields, improved efficiency and automation of normally manual processes within the laboratory is an active area of research. These various endeavors result in an ever changing landscape in the IVF laboratory.
This chapter focuses on various cryoprotectants and their application in preserving gametes and embryos. Penetrating cryoprotectants provide buffering against salt-induced stress by acting as solvent, reducing the solute concentration in the remaining water fraction inside the cell until the system is cooled to a low enough temperature. Various penetrating cryoprotectants have been used successfully in in vitro fertilization (IVF) to preserve sperm, embryos, and oocytes. Non-penetrating cryoprotectants are often included in media used for warming/thawing of cells to avoid osmotic shock. Most toxicity concerns exist with penetrating cryoprotectants rather than the non-penetrating cryoprotectants. In regard to reproductive biology and assessing suitability/toxicity of a cryoprotective agent, certain endpoint measures are commonly assessed. Cryoprotectants serve to prevent damage induced by the cooling/freezing process, in part by reducing ice crystal formation and by reducing stress resulting from osmotic shock.