Cryopreservation of Adult Mouse Testicular Germ Cells

Adult mouse testicular germ cells were cryopreserved,and the cell recovery rates were examined by trypan blue exclusion staining after thawing.The freezing solutions were DMEM/10% NBS supplemented with 5%,10%,15%,20% and 25% cryoprotectants dimethyl sulphoxide(DMSO),propanediol(PG),ethylene glycol(EG) or glycerol(G),respectively.The cell recoveries of DMSO at concentrations of 5%,10%,15%,20%,and 25% were 88 5%,88 0%,80 0%,65 6%,and 51 3%,respectively.The cell recoveries of PG at concentrations of 5%,10%,15%,20%,and 25% were 87 2%,86 4%,79 0%,73 4%,and 40 1%,respectively.The cell recoveries of EG at concentrations of 5%,10%,15%,20%,and 25% were 86 6%,80 9%,60 8%,51 3%,and 30 0%,respectively.The cell recoveries of G at concentrations of 5%,10%,15%,20%,and 25% were 86 5%,86 3%,65 3%,36 0%,and 31 4%,respectively.When the concentrations of four cryoprotectants were 5% 10%,the cell recoveries all were highest,and showed significant difference as compared with those at concentration of 15%.There were no significant differences among the highest recoveries of four cryoprotectants.The minimum cell losses of four cryoprotectants after freezing/thawing were 4 8%,6 1%,6 7%,and 6 8%,respectively.The results suggest that DMSO,PG,EG and G all are suitable for cryopreservation of adult mouse germ cells and their favorable concentration all are at 5% 10%.The two step freezing procedure is one with high cell recovery rates for cryopreservation of adult mouse germ cells.