Trumpet shell, Charonia sauliae, is an endangered and valuable species, but its artificial propagation protocol has not been successfully established. To estimate the possibility of cryopreservation for larvae of C. s...Trumpet shell, Charonia sauliae, is an endangered and valuable species, but its artificial propagation protocol has not been successfully established. To estimate the possibility of cryopreservation for larvae of C. sauliae, which is a potential preparation for its artificial reproduction and further research, in this study a protocoi for the cryopreservation of veliger larvae of trumpet shell was optimized. Through a two-step cryopreservation procedure, four kinds of cryoprotectants （ethylene glycol, 1, 2-propanediol, dimethyl sulfoxide and glycerol） were employed at three concentrations （1.0, 1.5 and 2.0molL^-1） respectively and survival rates of larvae were determined after a storage of lh. The larvae frozen with these four cryoprotectants after 1 h storage were cultured, and then survival rates were determined at 24, 72 and 120 h after thawing. Dimethyl sulfoxide at a concentration of 1.5 molL^-1 showed the best protective effect in all experiments （p〈0.05）. And survival rates of larvae frozen with dimethyl sulfoxide were determined after 1, 7 and 15 d of storage. The survival rates of larvae frozen with 1.5 molL^-1 dimethyl sulfoxide after 1 h, 1 d, 7 d and 15 d of storage were 80.77% ±7.51%, 80.34%±11.28%, 83.10%±9.14% and 77.23%±6.22% respectively. No significant differences in survival rates of larvae frozen with dimethyl sulfoxide were observed after various storage periods （p〉0.05）.展开更多
Objective: The aim of the study was to observe and compare the effects of cryopreservation and thawing meth- ods on rat ovarian tissues. Methods: Twenty 5-6 weeks old SPF-SD female rats were randomly divided into two ...Objective: The aim of the study was to observe and compare the effects of cryopreservation and thawing meth- ods on rat ovarian tissues. Methods: Twenty 5-6 weeks old SPF-SD female rats were randomly divided into two groups, with ten rats in each group. Freshly isolated ovaries saved as a control (group 1: fresh ovaries) in formalin-fixed or vitrified immediately after dissection (group 2: vitrified ovaries). Ovaries in vitrified group were processed into thin slices then cryo- preserved, stored in liquid nitrogen for 21 days, rapidly thawed and grossly examined. All of the collected ovaries underwent hematoxylin and eosin-stained paraffin serial sections and observed the microscopic evaluation in vitrified ovaries. Results: Grossly the vitrified ovaries turned pale color and the size was same as before freeze. The vitrified ovarian tissue had normal anatomical structures of cortex and medulla under the microscope and had no difference with the fresh control ovarian tis- sue. The number and distribution of the follicles were similar with the fresh ovarian tissue, but had smaller size and the gap between oocyte and the surrounding granulosa cells was increased. Few ooctyes were in irregular appearance however the morphology of follicular cells did not give a different appearance as compared to the fresh control ovarian tissue. Conclusion: Cryopreservation of ovarian tissues by vitrification method has some detrimental effect on the morphology of follicles but does not induce negative impact on the number, density and survival of the primordial ovarian follicles. However the whole follicle anatomical structures also had no significant changes.展开更多
文摘Trumpet shell, Charonia sauliae, is an endangered and valuable species, but its artificial propagation protocol has not been successfully established. To estimate the possibility of cryopreservation for larvae of C. sauliae, which is a potential preparation for its artificial reproduction and further research, in this study a protocoi for the cryopreservation of veliger larvae of trumpet shell was optimized. Through a two-step cryopreservation procedure, four kinds of cryoprotectants （ethylene glycol, 1, 2-propanediol, dimethyl sulfoxide and glycerol） were employed at three concentrations （1.0, 1.5 and 2.0molL^-1） respectively and survival rates of larvae were determined after a storage of lh. The larvae frozen with these four cryoprotectants after 1 h storage were cultured, and then survival rates were determined at 24, 72 and 120 h after thawing. Dimethyl sulfoxide at a concentration of 1.5 molL^-1 showed the best protective effect in all experiments （p〈0.05）. And survival rates of larvae frozen with dimethyl sulfoxide were determined after 1, 7 and 15 d of storage. The survival rates of larvae frozen with 1.5 molL^-1 dimethyl sulfoxide after 1 h, 1 d, 7 d and 15 d of storage were 80.77% ±7.51%, 80.34%±11.28%, 83.10%±9.14% and 77.23%±6.22% respectively. No significant differences in survival rates of larvae frozen with dimethyl sulfoxide were observed after various storage periods （p〉0.05）.
文摘Objective: The aim of the study was to observe and compare the effects of cryopreservation and thawing meth- ods on rat ovarian tissues. Methods: Twenty 5-6 weeks old SPF-SD female rats were randomly divided into two groups, with ten rats in each group. Freshly isolated ovaries saved as a control (group 1: fresh ovaries) in formalin-fixed or vitrified immediately after dissection (group 2: vitrified ovaries). Ovaries in vitrified group were processed into thin slices then cryo- preserved, stored in liquid nitrogen for 21 days, rapidly thawed and grossly examined. All of the collected ovaries underwent hematoxylin and eosin-stained paraffin serial sections and observed the microscopic evaluation in vitrified ovaries. Results: Grossly the vitrified ovaries turned pale color and the size was same as before freeze. The vitrified ovarian tissue had normal anatomical structures of cortex and medulla under the microscope and had no difference with the fresh control ovarian tis- sue. The number and distribution of the follicles were similar with the fresh ovarian tissue, but had smaller size and the gap between oocyte and the surrounding granulosa cells was increased. Few ooctyes were in irregular appearance however the morphology of follicular cells did not give a different appearance as compared to the fresh control ovarian tissue. Conclusion: Cryopreservation of ovarian tissues by vitrification method has some detrimental effect on the morphology of follicles but does not induce negative impact on the number, density and survival of the primordial ovarian follicles. However the whole follicle anatomical structures also had no significant changes.
