Novel Approaches to the Cryopreservation of Human Spermatozoa: History and Development of the Spermatozoa Vitrification Technology

Cryobiology is very intensively applied in reproductive and veterinary medicine for preservation of gametes, embryos and reproductive tissues. Sub-zero temperatures combined with appropriate cryoprotective agents preserve the physiological and reproductive functions of the cells making long-term storage possible without loss of viability. With the use of cryoprotective agents it has become possible to develop cryopreservation techniques, such as the slow conventional freezing and vitrification that are in use in the present times. In slow controlled-rate conventional freezing extracellular ice crystals are formed whereas in vitrification no ice crystals are formed. Glass formation is compatible with the survival of the cell and the preservation of its intracellular structures provided the type(s) and concentrations of cryoprotectant used are not chemo- or osmotoxic. However, irrespective of the type of cooling method employed the cryosurvival of cells and tissues is influenced by the size and maturity of cells, amounts of intracellular water, quality and quantity of intracellular lipids, type of cells, their function and morphology. The intracellular milieu of cryopreserved cells and tissues remain less understood. The application of nanotechnology may help reveal and help advance our knowledge of the cryobiological principles involved in cryosurvival. At this moment the methods of cryopreservation that merit further investigation are vitrification and lyophilization. Vitrification is cheap if reagents are prepared in-house and the procedure can be performed rapidly. It has been successfully applied for gametes and embryos (of different stages of development), and reproductive cells/tissues, somatic cells and stem cells. However, vitrification is more demanding technically and requires operation and storage at sub-zero temperatures. On the other hand lyophilization deserves further investigation because it is a cheaper form of cryopreservation that may enable cryostorage at less demanding temperatures of 4°C and may even allow transport at ambient temperature. These possibilities are explored in this review.