Comparison of the effects of three cryoprotectants on the cryopreservation of mouse subcutaneous tissue under different conditions

Abstract

The subcutaneous tissue of animals contains different cell types, and different cells have different requirements for cryopreservation. This establishes obstacles that need to be overcome in the clinical application of tissue preservation. In the present study, the effects of different freezing rates and various concentrations of cryoprotectants on the cryopreservation of subcutaneous tissue of mice were compared, and these results provided basic research data that can be used to explore the optimal cryopreservation method for tissue. The effects of three cryoprotectants, dimethyl sulfoxide, glycerinum and 1,2-propanediol, and their concentrations on the cryopreservation of subcutaneous tissue of mice were compared with slow and rapid freezing rates. The results revealed that under various cryopreservation conditions, the percentage of fibroblasts that grow from the tissue following slow cryopreservation (19.8%) was significantly higher than that following rapid freezing (6.7%) at osmotic equilibrium for 10-20 min (P<0.05). After 19 days of culture, under the conditions of slow freezing, with 10, 20 and 30% glycerinum as a cryoprotectant, respectively, fibroblasts grew from 26.0, 16.7 and 16.7% of the tissues, respectively. No fibroblasts were indicated in the tissue mass cultured in any other tissue blocks treated with cryopreservation solutions. Under the condition of rapid freezing, fibroblasts grew from 6.7 and 6.7% tissue blocks of 20% DMSO and 10% glycerinum, respectively, following 19 days of culture. No fibroblasts were identified in the tissue mass cultured in the other tissue blocks treated with cryopreservation solutions, and no fibroblasts were identified in the tissue blocks without osmotic balance before freezing.

Evaluating the Effects of Cryopreservation on the Viability and Gene Expression of Porcine-Ear-Skin Fibroblasts

“…For instance, Naaldijk et al compared the effects of different machine-based and manual cryopreservation methods on the post-thaw survival and proliferative capacity of human fibroblasts, and suggested that the freezing-container method was the most effective solution [ 4 ]. Wang et al revealed that mouse ( Mus musculus ) fibroblasts’ post-thaw recovery rate under slow cryopreservation was significantly higher than that under rapid freezing [ 16 ]. Costa et al indicated that both direct vitrification in cryovials and solid-surface vitrification were effective in the preservation of agouti ( Dasyprocta leporina Linnaeus ) fibroblasts, but led to an increase in the number of perinuclear halos [ 17 ].…”