Cryopreservation and microfluidics: a focus on the oocyte.

Abstract

Cryopreservation of gametes and embryos has played a critical role in successful assisted reproductive technologies in rodents, domestic farm species, endangered species and humans. With improved success, and changing needs, the utility of gamete or embryo cryopreservation has escalated. In this review we address some of the foundational history of mammalian cryobiology, species-specific utilities, fundamental understandings of cryoprotectant agents and their use in slow-rate freezing and vitrification, and expand on the recent success and uses of oocyte vitrification and warming. In the area of female gamete cryopreservation, emphasis will be placed on not just cell survival, but also perceived and measured affects of cryopreservation on intracellular structures and functions that affect subsequent completion of meiosis with chromatin segregation fidelity, normal fertilisation and embryonic developmental competence. We compare and contrast data from cow, mouse and humans with a focus on using species-comparative developmental biology to guide future studies for improving methodologies for all species. The application of the relatively new technology microfluidics is discussed in relation to moving gradually (i.e. changing the solution over cells in an automated fashion) compared with the stepwise manual movement of cells through changing solution currently used. This use of microfluidics to change the way cells are exposed to cryoprotectant agents can provide new insights into the effects of osmotic stress and cellular strain rates previously unappreciated, precise methods of computational and biological data acquisition and appreciation of morphometric changes to cellular structure in response to different osmotic stresses and strain rates achieved with varying cryoprotectant exposures. Collectively, these devices and methodologies provide a means of achieving incremental improvement of oocyte and zygote cryopreservation with normalised and improved developmental competence. Finally, we look to the past and the future to acknowledge the accomplishment of leaders in the field of mammalian gamete and embryo cryobiology, their inspirational works, their tireless dissemination of information and the potential of new technologies in bioengineering to improve the efficiency and safety of gamete and embryo cryopreservation.