This study was performed to find out the optimal kind of additive and concentration of ethylene glycol (EG) as the cryoprotectant on the D-shaped larvae of arkshell, Scapharca broughtonii. The larvae in straws was carried in the programed freezer set ...
This study was performed to find out the optimal kind of additive and concentration of ethylene glycol (EG) as the cryoprotectant on the D-shaped larvae of arkshell, Scapharca broughtonii. The larvae in straws was carried in the programed freezer set up at 0℃, frozen to -12℃ by the freezing rate of -1℃/min, held for 10 minutes, seeded at -12℃, finally refrozen to -35℃ by the freezing rate of -1℃/min and then directly plunged into liquid nitrogen. The survival of larvae frozen in 1.0 M and 2.0 M EG added with 0.5 M sucrose were high (50.5 ± 1.3% and 51.9 ± 1.7%, respectively) in the early D-shaped larvae cryopreserved in 1 M and 2 M EG diluted with 0.2 M and 0.5 M fructose, glucose and sucrose. In the late D-shaped larvae were cryopreserved according to five concentrations of EG added with 0.5 M sucrose, the survival of larvae frozen in 2.0 M EG was the highest as 51.9 ± 1.7%. The morphological differences in cells between unfrozen and frozen-thawed D-shaped larvae were not found by the scanning and transmission electron microscopy.