ISSN : 1225-3480
피조개 D형 유생을 냉동보존하기 위한 적절한 첨가제 종류와 농도 및 동해방지제로 적용한 EG의 농도에 대한 냉동보존 효과실험을 실시하였다. 유생을 0℃로 설정된 프로그램 냉동기로 옮겨 -1℃/분의 냉동률로 -12℃까지 냉동한 후 10분 동안 유지한 상태에서 식빙하고 -35℃까지 재냉동한 다음, 즉시 액체질소통에 저장하였다. 1.0 M과 2.0 M EG을 동해방지제로 사용하여 첨가제별로 D형 유생을 냉동한 결과, 0.5 M sucrose에서 생존율이 각각 50.5 ± 1.3%, 51.9 ± 1.7%로 가장 좋았다. 0.5 M sucrose를 첨가한 EG의 농도별 냉동에서는 2.0 M이 생존율 51.9 ± 1.7%로 가장 높았다. 주사 및 투과전자현미경을 이용하여 유생을 관찰한 결과, 냉동 전, 후 유생의 외부형태 및 세포내 구조의 차이가 인정되지 않았다.
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.
