The Effects of Cryoprotectant Combinations and Cryopreservation Durations on the Survival and Fecundity of Wild-Type Caenorhabditis Elegans

Abstract

Cryopreservation is the process of storing biological structures in low temperatures for future utilization. Different cryoprotectants and combinations are used to improve the quality of the preserved specimen upon thawing. However, there is no standardized methodology yet for the model organism Caenorhabditis elegans. The aim of this research was to determine the effects of different combinations of cryopreserving compounds and different lengths of cryopreservation on the survivorship and fecundity of the N2 C. elegans strain. Multiple treatments were studied, each consisting of varying combinations of cryoprotectants, specifically 50% glycerol and 50% sucrose, 75% glycerol and 25% sucrose, 25% glycerol and 75% sucrose, 50% glycerol and 50% trehalose, 75% glycerol and 25% trehalose, as well as 25% glycerol and 75% trehalose, all in M9 Buffer. Observations were conducted at three different durations of cryopreservation (5, 10, and 15 days) for each of the cryopreservation setups. It was determined that different CPA combinations and durations of cryopreservation significantly affected the fecundity of the C. elegans samples, but not their survival rate. In conclusion, the 25% glycerol & 75% sucrose treatment is the best CPA combination among all the treatments studied and the 15-day cryopreservation yielded the greatest number of progeny. However, further study is necessary to verify the effects of cryopreservation duration on the fecundity of C. elegans.