Authors : Trinidad Leon-Quinto, Sisco Manas and Arturo Serna
Abstract: Cryobanked somatic cells may be used when required for wide-ranging applications such as advanced veterinary reproductive biotechnologies, genetic resources conservation or studies at genetic level, among others. In populations where gametes are difficult to obtain such as threatened species, such cells acquire special relevance because they could be later reintroduced into the breeding pool by inducing pluripotency and/or nuclear transfer. Since, knowledge of specific cell response to cryopreservation process is required, the physiological sensitivity to cryoinjury of skin cells from the critically endangered European mink was evaluated in this research and for the first time. After thawing, the cell viability and functionality was investigated by analyzing their membrane integrity, metabolic activity, glycosaminoglycan production and proliferative capability. Freezing media included the permeating cryoprotectant dimethyl sulfoxide either at 5 or 10%. These two concentrations were investigated alone or along with the non-permeating cryoprotectant sucrose at 0.1 or 0.2 M, testing six different freezing solutions. Dimethylsulfoxide at 10% resulted in thawed cells exhibiting proliferative capabilities similar to those of fresh-unfrozen cells with mean values around 80, 90 and 85% for viability, metabolic activity and GAG synthesis, respectively. Dimethylsulfoxide alone at 5% resulted in a significantly decreased protection. However, the combination of 5%-dimethyl sulfoxide and sucrose (both 0.1 as 0.2 M) yielded similar results to those obtained with 10%-dimethylsulfoxide. The findings are relevant to preserve alive the genetic patrimony of the European mink as well as to provide properly cryobanked cells for a wide variety of applications. The results could be also useful for other mammals.
Trinidad Leon-Quinto, Sisco Manas and Arturo Serna, 2013. Cryobanking of Skin Cells from the Critically Endangered European Mink (Mustela lutreola). Journal of Animal and Veterinary Advances, 12: 1665-1672.