Objective To study the efficacy of two vitrification solutions for mouse morulae Methods Good morulaes of NIH mice were collected and used to test toxicity of the vitrification solutions EDS40 （40% ethylene glycol, 1...Objective To study the efficacy of two vitrification solutions for mouse morulae Methods Good morulaes of NIH mice were collected and used to test toxicity of the vitrification solutions EDS40 （40% ethylene glycol, 18% dextran and 0.5 tool sucrose） and EFS40 （40% ethylene glycol, 18% ficoll and 0.5 tool sucrose）. Fine vitrified morulae were packaged in 0.25 mL plastic straws and immersed into liquid nitrogen and cryopreserved for about 2-3 months. Then the straws were heated rapidly, washed in Ham＇s F12 medium and cultured. The viability was determined by morphology and blastocyst formation after being cultured for 48 h. Some embryos were transplanted to recipients after being cultured for 12-14 h. The number of pregnant recipients and young born was counted and analyzed by Chi-squared test. Results The toxicity of EDS40 solution was significantly lower than that of EFS40 （P〈0.05） and the number of embryos developed to the blastocysts after vitrification in EDS40 was significantly higher than in EFS40 （P〈0.05）. The number of zona pellucida and the number of pregnancy and birth integrated after vitrification cryopreservation had no significant difference between EDS40 and EFS40 （P〉0.05）. However, the embryo fineness rates after vitrification in EDS40 was significantly better than in EFS40 （P〈0.01）. Conclusion EDS40 solution has less toxicity and better cryoprotect effect on embryos than EFS 40.展开更多
Objective: To assess the developing potentiality of mouse morula produced in vitro or in the after vitrification and to evaluate the effect of one-step and two-step vitrification methods. Method: Mouse morula produced...Objective: To assess the developing potentiality of mouse morula produced in vitro or in the after vitrification and to evaluate the effect of one-step and two-step vitrification methods. Method: Mouse morula produced in for and in the were vitrified in the solution containing ethylene glycol, Ficoll and sucrose (EFS solution) with one-step and two-step methods. The developing potential and status of the pellucid zona in vitified mouse morula were assessed. Results: The percentages of morula developed into blastocyst stage were 81. 8% and 82.4%, 97. 3% and 98.4%, respectively, after one-step and two-step exposure of in vitro morula or in vivo morula to EFS solution alone, which did not show significant difference compared to their controls (P > 0. 05). The percentage of in vitro morula developed into blastocyst vitrified by onestep method was significantly lower than that by two-step method and coned (P < 0.05, 70.6% vs 81 .3%; 70.6% vs 83 .6%, respectively). However, there was no significant difference between blastocyst rates of in vivo morula vitrified by one-step and two-step methods (P>0.05, 93. 1% us 95.7%). No rupture of pellucid zona was observed in all thawed morula after one-step and two-step vitrification, irrespective of in vitro morula or in vivo morula. Conclusion: Morula produced in vitro and in vivo after vitrification may maintain high survival rate and developing potential. Two-step vitrification method with EFS solution is suitable for in vitro morula, which can improve the developing potential of in vitro morula. Onestep and two-step vitrification method have no detrimennd effect on the developing potential of in vivo morula.展开更多
[Objectives]This study was conducted to explore the effects of embryos at different developmental stages on the bisection effect of embryos,improve the efficiency of bovine embryo bisection,and facilitate the applicat...[Objectives]This study was conducted to explore the effects of embryos at different developmental stages on the bisection effect of embryos,improve the efficiency of bovine embryo bisection,and facilitate the application of embryo bisection technology in cattle breeding.[Methods]The effects of two different bisection solutions on the bisection of morulae and blastocysts in vitro were explored.The morulae and blastocysts produced in vitro from cattle that developed to the 6th to 8th d were bisected by hands,and demi-embryos were cultured in vitro.Their development was observed.[Results]Morulae were bisected in PBS solution and PBS+0.2 mol/L sucrose,and the success rates of bisection were 50%and 95.2%,respectively.The success rate of bisecting morulae in PBS+0.2 mol/L sucrose was significantly higher than that in PBS(P<0.05),while the development rate of the bisected demi-embryos had no significant difference between the two(53.3%,52.4%)(P>0.05).The success rates of blastocyst bisection in PBS solution and PBS+0.2 mol/L sucrose were 51.6%and 95.1%,respectively.The success rate of blastocyst bisection in PBS+0.2 mol/L sucrose was significantly higher than that in PBS(P<0.05),while the development rate of the bisected demi-embryos had no significant difference between the two(50.0%,56.4%)(P>0.05).[Conclusions]There were no significant differences between the success rates of bisecting bovine morulae and blastocysts in PBS+0.2 mol/L sucrose,which were both significantly better than those in pure PBS bisection solution,proving that PBS+0.2 mol/L sucrose bisection solution is suitable for bovine embryo bisection.展开更多
The capacities of phagocytes of subpopulation P1 (F) and morula cells (MC) of holothurian Eupentacta fraudatrix to modulate apoptosis of each other as well as cytokine-dependent mechanisms and hormonal regulation of t...The capacities of phagocytes of subpopulation P1 (F) and morula cells (MC) of holothurian Eupentacta fraudatrix to modulate apoptosis of each other as well as cytokine-dependent mechanisms and hormonal regulation of these cells’s interaction were studied. The 18-h treatment of F with supernatants, obtained after centrifugation of MC preincubated for 3 h with phosphate buffered saline (PSB) at the temperature of 22℃?(SMC3) resulted in a significant growth of apoptosis level. A 30-min incubation of F with supernatants of MC, preincubated for 24 h (SMC24), on the contrary, reduced the apoptosis level and increased the level of interleukine-1α (IL-1α)-like substances, and 24-h incubation did not influence apoptosis and reduced level of IL-1α-like substances. Thus, proapoptotic effects of MC’s supernatants in F inversely depended on time of their preincubation with PSB and directly on time of incubation with F. Additionally, this effect was opposite to variations in the level of IL-1α-like substances. The level of apoptosis declined after 30 min of incubation but elevated after 24 h at the inverse treatment of MC with supernatant, obtained after preincubation of F during 24 h (SF24). The level of IL-1α-like substances dropped after 30 min and insignificantly decreased after 24 h. Hence, SF24 proapoptotic effect directly depended on time of incubation with MC and did not correspond to variations in the level of IL-1α-like substances. 100 μM dexamethasone stimulated apoptosis in F and MC in an inverse time-dependent manner during 24-h preincubation, and supernatants of cell suspensions obtained after such preincubations, stimulated apoptosis and reduced the IL-1α-like substances level in target cells at both types of interaction. IL-1α-like substances are?supposed to be mediators for MC’s effects in F, but not for F’s action on MC. In holothurians, steroid hormones apparently may participate in the regulation of the immune response and cell cooperation.展开更多
Since the first report on the successful deep cryopreservation of mam-malian embryos in 1972,slow progressing cooling rate has been employ-ed in conventional embryos-freezing techniques;while more recent studieson fre...Since the first report on the successful deep cryopreservation of mam-malian embryos in 1972,slow progressing cooling rate has been employ-ed in conventional embryos-freezing techniques;while more recent studieson freezing preimplantation embryos have focussed on the simplificationof cooling and thawing procedures and improvement of viability of embr-yos.Dimethylsulfoxide (DMSO)has been used as an effective cryoprotec-tant for freezing human and other mammalian embryos.It has been foundthat glycerol and other alcohols are effective to protect embryos fromcryoinjury展开更多
基金This study was supported by Chongqing Medical Foundation of Science and Technology.
文摘Objective To study the efficacy of two vitrification solutions for mouse morulae Methods Good morulaes of NIH mice were collected and used to test toxicity of the vitrification solutions EDS40 （40% ethylene glycol, 18% dextran and 0.5 tool sucrose） and EFS40 （40% ethylene glycol, 18% ficoll and 0.5 tool sucrose）. Fine vitrified morulae were packaged in 0.25 mL plastic straws and immersed into liquid nitrogen and cryopreserved for about 2-3 months. Then the straws were heated rapidly, washed in Ham＇s F12 medium and cultured. The viability was determined by morphology and blastocyst formation after being cultured for 48 h. Some embryos were transplanted to recipients after being cultured for 12-14 h. The number of pregnant recipients and young born was counted and analyzed by Chi-squared test. Results The toxicity of EDS40 solution was significantly lower than that of EFS40 （P〈0.05） and the number of embryos developed to the blastocysts after vitrification in EDS40 was significantly higher than in EFS40 （P〈0.05）. The number of zona pellucida and the number of pregnancy and birth integrated after vitrification cryopreservation had no significant difference between EDS40 and EFS40 （P〉0.05）. However, the embryo fineness rates after vitrification in EDS40 was significantly better than in EFS40 （P〈0.01）. Conclusion EDS40 solution has less toxicity and better cryoprotect effect on embryos than EFS 40.
文摘Objective: To assess the developing potentiality of mouse morula produced in vitro or in the after vitrification and to evaluate the effect of one-step and two-step vitrification methods. Method: Mouse morula produced in for and in the were vitrified in the solution containing ethylene glycol, Ficoll and sucrose (EFS solution) with one-step and two-step methods. The developing potential and status of the pellucid zona in vitified mouse morula were assessed. Results: The percentages of morula developed into blastocyst stage were 81. 8% and 82.4%, 97. 3% and 98.4%, respectively, after one-step and two-step exposure of in vitro morula or in vivo morula to EFS solution alone, which did not show significant difference compared to their controls (P > 0. 05). The percentage of in vitro morula developed into blastocyst vitrified by onestep method was significantly lower than that by two-step method and coned (P < 0.05, 70.6% vs 81 .3%; 70.6% vs 83 .6%, respectively). However, there was no significant difference between blastocyst rates of in vivo morula vitrified by one-step and two-step methods (P>0.05, 93. 1% us 95.7%). No rupture of pellucid zona was observed in all thawed morula after one-step and two-step vitrification, irrespective of in vitro morula or in vivo morula. Conclusion: Morula produced in vitro and in vivo after vitrification may maintain high survival rate and developing potential. Two-step vitrification method with EFS solution is suitable for in vitro morula, which can improve the developing potential of in vitro morula. Onestep and two-step vitrification method have no detrimennd effect on the developing potential of in vivo morula.
基金Qiqihar Agricultural Research Project(NYGG-201904)China Agriculture Research System of MOF and MRRA.
文摘[Objectives]This study was conducted to explore the effects of embryos at different developmental stages on the bisection effect of embryos,improve the efficiency of bovine embryo bisection,and facilitate the application of embryo bisection technology in cattle breeding.[Methods]The effects of two different bisection solutions on the bisection of morulae and blastocysts in vitro were explored.The morulae and blastocysts produced in vitro from cattle that developed to the 6th to 8th d were bisected by hands,and demi-embryos were cultured in vitro.Their development was observed.[Results]Morulae were bisected in PBS solution and PBS+0.2 mol/L sucrose,and the success rates of bisection were 50%and 95.2%,respectively.The success rate of bisecting morulae in PBS+0.2 mol/L sucrose was significantly higher than that in PBS(P<0.05),while the development rate of the bisected demi-embryos had no significant difference between the two(53.3%,52.4%)(P>0.05).The success rates of blastocyst bisection in PBS solution and PBS+0.2 mol/L sucrose were 51.6%and 95.1%,respectively.The success rate of blastocyst bisection in PBS+0.2 mol/L sucrose was significantly higher than that in PBS(P<0.05),while the development rate of the bisected demi-embryos had no significant difference between the two(50.0%,56.4%)(P>0.05).[Conclusions]There were no significant differences between the success rates of bisecting bovine morulae and blastocysts in PBS+0.2 mol/L sucrose,which were both significantly better than those in pure PBS bisection solution,proving that PBS+0.2 mol/L sucrose bisection solution is suitable for bovine embryo bisection.
文摘The capacities of phagocytes of subpopulation P1 (F) and morula cells (MC) of holothurian Eupentacta fraudatrix to modulate apoptosis of each other as well as cytokine-dependent mechanisms and hormonal regulation of these cells’s interaction were studied. The 18-h treatment of F with supernatants, obtained after centrifugation of MC preincubated for 3 h with phosphate buffered saline (PSB) at the temperature of 22℃?(SMC3) resulted in a significant growth of apoptosis level. A 30-min incubation of F with supernatants of MC, preincubated for 24 h (SMC24), on the contrary, reduced the apoptosis level and increased the level of interleukine-1α (IL-1α)-like substances, and 24-h incubation did not influence apoptosis and reduced level of IL-1α-like substances. Thus, proapoptotic effects of MC’s supernatants in F inversely depended on time of their preincubation with PSB and directly on time of incubation with F. Additionally, this effect was opposite to variations in the level of IL-1α-like substances. The level of apoptosis declined after 30 min of incubation but elevated after 24 h at the inverse treatment of MC with supernatant, obtained after preincubation of F during 24 h (SF24). The level of IL-1α-like substances dropped after 30 min and insignificantly decreased after 24 h. Hence, SF24 proapoptotic effect directly depended on time of incubation with MC and did not correspond to variations in the level of IL-1α-like substances. 100 μM dexamethasone stimulated apoptosis in F and MC in an inverse time-dependent manner during 24-h preincubation, and supernatants of cell suspensions obtained after such preincubations, stimulated apoptosis and reduced the IL-1α-like substances level in target cells at both types of interaction. IL-1α-like substances are?supposed to be mediators for MC’s effects in F, but not for F’s action on MC. In holothurians, steroid hormones apparently may participate in the regulation of the immune response and cell cooperation.
文摘Since the first report on the successful deep cryopreservation of mam-malian embryos in 1972,slow progressing cooling rate has been employ-ed in conventional embryos-freezing techniques;while more recent studieson freezing preimplantation embryos have focussed on the simplificationof cooling and thawing procedures and improvement of viability of embr-yos.Dimethylsulfoxide (DMSO)has been used as an effective cryoprotec-tant for freezing human and other mammalian embryos.It has been foundthat glycerol and other alcohols are effective to protect embryos fromcryoinjury
