Objective To investigate the viability and maturation of frozen-thawed human immature oocytes exposed to different temperature of vitrification and warming solutions. Methods The immature oocytes in germinal vesicle ...Objective To investigate the viability and maturation of frozen-thawed human immature oocytes exposed to different temperature of vitrification and warming solutions. Methods The immature oocytes in germinal vesicle （GV） and matephase I （MI ） stages were collected from our ICSI patients and exposed to different temperature of vitrification and warming solutions before frozen in the freezing/thawing procedures. The different temperature groups were as follows： Group A, equilibration solution at 37℃, vitrification solution at room temperature, warming solution at 37℃; Group B, both vitrification and warming solution at room temperature; Group C, both vitrification and warming solutions at 37℃; Group D, the frozen-thawed oocytes and the fresh oocytes were cultured for in vitro maturation. The survival rate and maturation rate were compared among groups. The oocytes were examined using immunofluorescent stainingand confocal microscopy to check the spindle configuration and chromosome arrangement. Results The survival rates and MII rates of GV stage oocytes in groups A, B, C were 100%（15/15）, 81.3%（13/16）, 68.8%（11/16） and 33.3%（2/6）, 83.3%（10/12）,72.7%（8/11）, respectively. The survival rate of group C was significantly lower than that of the control （P〈0.05）. The normal spindle and chromosome configuration were only observed in group B, with the rates of 20% （2/20） and 10% （1/10）, respectively. The survival rates of MI stage in groups A, B, C were 71.4%（10/14）, 100% （12/12） and 83.3%（10/12）, no significantly difference from that of the contro1（100%, 14/14）. The MII rates of MI stage in groups A, B and C were 0%（0/14）, 66.7%（8/12） and 80% （8/10）, respectively. The MⅡ rate in group A was significantly lower than that in other groups （P〈0.01）. Only one oocyte in group C was found with normal spindle and chromosome configurations. Conclusion The appropriate operation temperature of vitrification and warming solutions can improve the outcomes of the vitrified-thawed human immature oocytes.展开更多
Aim: To evaluate the fertilization competence of spermatozoa from ejaculates and testicle when the oocytes were matured in vitro following intracytoplasmic sperm injection (ICSI). Methods: Fifty-six completed cycles i...Aim: To evaluate the fertilization competence of spermatozoa from ejaculates and testicle when the oocytes were matured in vitro following intracytoplasmic sperm injection (ICSI). Methods: Fifty-six completed cycles in 46 women with polycystic ovarian syndrome were grouped according to the semen parameters of their male partners. Group 1 was 47 cycles that presented motile and normal morphology spermatozoa in ejaculates and Group 2 was the other nine cycles where male partners were diagnosed as obstructive azoospermia and spermatozoa could only be found in testicular tissue fragment. All female patients received minimal stimulation with gonadotropin. Immature oocytes were matured in vitro and inseminated by ICSI. The spermatozoa from testes were retrieved by testicular fine needle aspiration. Results: A total of 449 and 78 immature oocytes were collected and cultured for 48 hours, 75.5 % (339/449) and 84.6 % (66/78) oocytes were matured in Groups 1 and 2, respectively. The percentage of oocytes achieving normal fertilization was significantly higher in Group 1 than that in Group 2 (72.9 % vs. 54.5 %, P < 0.05). There were no significant differences in the rates of oocytes cleavage and clinical pregnancies in these two groups [87.4 % (216/247) vs. 88.9 % (32/36); 21.3 % (10/47) vs. 44.4 % (4/9)]. A total of 15 babies in the two groups were healthy delivered at term. Conclusion: It appears that IVM combined with ICSI using testicular spermatozoa can produce healthy infants, while the normal fertilization rate of in vitro matured oocytes after ICSI using testicular spermatozoa was significantly lower than using the ejaculated spermatozoa.展开更多
文摘Objective To investigate the viability and maturation of frozen-thawed human immature oocytes exposed to different temperature of vitrification and warming solutions. Methods The immature oocytes in germinal vesicle （GV） and matephase I （MI ） stages were collected from our ICSI patients and exposed to different temperature of vitrification and warming solutions before frozen in the freezing/thawing procedures. The different temperature groups were as follows： Group A, equilibration solution at 37℃, vitrification solution at room temperature, warming solution at 37℃; Group B, both vitrification and warming solution at room temperature; Group C, both vitrification and warming solutions at 37℃; Group D, the frozen-thawed oocytes and the fresh oocytes were cultured for in vitro maturation. The survival rate and maturation rate were compared among groups. The oocytes were examined using immunofluorescent stainingand confocal microscopy to check the spindle configuration and chromosome arrangement. Results The survival rates and MII rates of GV stage oocytes in groups A, B, C were 100%（15/15）, 81.3%（13/16）, 68.8%（11/16） and 33.3%（2/6）, 83.3%（10/12）,72.7%（8/11）, respectively. The survival rate of group C was significantly lower than that of the control （P〈0.05）. The normal spindle and chromosome configuration were only observed in group B, with the rates of 20% （2/20） and 10% （1/10）, respectively. The survival rates of MI stage in groups A, B, C were 71.4%（10/14）, 100% （12/12） and 83.3%（10/12）, no significantly difference from that of the contro1（100%, 14/14）. The MII rates of MI stage in groups A, B and C were 0%（0/14）, 66.7%（8/12） and 80% （8/10）, respectively. The MⅡ rate in group A was significantly lower than that in other groups （P〈0.01）. Only one oocyte in group C was found with normal spindle and chromosome configurations. Conclusion The appropriate operation temperature of vitrification and warming solutions can improve the outcomes of the vitrified-thawed human immature oocytes.
文摘Aim: To evaluate the fertilization competence of spermatozoa from ejaculates and testicle when the oocytes were matured in vitro following intracytoplasmic sperm injection (ICSI). Methods: Fifty-six completed cycles in 46 women with polycystic ovarian syndrome were grouped according to the semen parameters of their male partners. Group 1 was 47 cycles that presented motile and normal morphology spermatozoa in ejaculates and Group 2 was the other nine cycles where male partners were diagnosed as obstructive azoospermia and spermatozoa could only be found in testicular tissue fragment. All female patients received minimal stimulation with gonadotropin. Immature oocytes were matured in vitro and inseminated by ICSI. The spermatozoa from testes were retrieved by testicular fine needle aspiration. Results: A total of 449 and 78 immature oocytes were collected and cultured for 48 hours, 75.5 % (339/449) and 84.6 % (66/78) oocytes were matured in Groups 1 and 2, respectively. The percentage of oocytes achieving normal fertilization was significantly higher in Group 1 than that in Group 2 (72.9 % vs. 54.5 %, P < 0.05). There were no significant differences in the rates of oocytes cleavage and clinical pregnancies in these two groups [87.4 % (216/247) vs. 88.9 % (32/36); 21.3 % (10/47) vs. 44.4 % (4/9)]. A total of 15 babies in the two groups were healthy delivered at term. Conclusion: It appears that IVM combined with ICSI using testicular spermatozoa can produce healthy infants, while the normal fertilization rate of in vitro matured oocytes after ICSI using testicular spermatozoa was significantly lower than using the ejaculated spermatozoa.
