Cryopreservation of the cells from buffalo seminiferous epithelia.

The aim of this study was to explore the effects of cryopreservation with different cryoprotectants and concentration for cells from buffalo seminiferous epithelia.The cells from 3-to 5-month-old buffalo seminiferous epithelia were resuspended in DMEM medium supplemented with 10% fetal bovine serum(FBS) and varying concentrations of dimethylsulfoxide(DMSO),glycerol(G),propylene glycol(PG) and ethylene glycol(EG),and these cells survival rates were assessed by trypan blue exclusion staining after freezing.In cryoprotective medium supplemented with 0%,5%,10%,15%,20% DMSO,the cells frozen/thawed in medium containing 10% DMSO had a significantly(P0.05) higher percentage of living cells compared to medium with other concentrations of DMSO.In cryoprotective medium supplemented with 0%,20%,25%,30%,35%,40% G,the cells frozen/thawed in medium containing 35% G had a significantly(P0.05) higher percentage of living cells compared to medium with other concentrations of G.In cryoprotective medium supplemented with 0%,5%,10%,15%,20%,25% PG,the cells frozen/thawed in medium containing 15%-25% PG all had a significantly(P0.05) higher percentage of living cells compared to medium with other concentration of PG,whereas the cell survival rate in medium containing 20% PG was the best after thawing.In cryoprotective medium supplemented with 0%,5%,10%,15%,20% EG,the cells frozen/thawed in medium containing 5%-20% EG all had a significantly(P0.05) higher percentage of living cells compared to medium with 0%EG,whereas the cell survival rate in medium contraining 10% EG was the best after thawing.The highest cell survival rates of the four cryoprotectants showed significant difference in medium among containing 10% DMSO or 35%G and containing 20% PG or 10% EG.The results suggested that it was a feasible approach of cryopreservation for cells from buffalo seminiferous epithelia by two-step freezing in DMEM medium supplemented with 10% FBS and 10% DMSO or 35% G,liquid nitrogen storing and 37℃ water bath thawing.