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• W2100084351 abstract "OBJECTIVE: After controlled ovarian hyperstimulation, most of recovered ova are mature at the metaphase II (MII: 85%) while others immature, being at either the metaphase I (MI: 4%) or the GV stage (11%). To study the nuclear and cytoplasmic competence of GV oocytes was aimed here.DESIGN: Experimental research, approved by the Ethical Comitee of the Instituto Universitario IVI, Law14/2003, Spain.MATERIALS AND METHODS: A total of 159 GV oocytes from stimulated cycles were donated to this research. Individual GV oocytes were matured in 50μL hTF medium for 2 days. Progression on nuclear maturation was assessed twice along the in vitro maturation (IVM) period; at IVM1 (20-24hrs) and at IVM2 (44-48hrs). At these check points, ova were checked for the presence of a nucleus (GV), absence of nucleus nor polar body (MI) or absence of nucleus, but presence of the 1st polar body in the perivitelline space (MII). To determine the cytoplasmic competence, 31 MII ova were activated artificially. Briefly, they were incubated serial in 5μM A23187 (5min) and 10μg/mL puromycin (6hrs). After rinsing, they were cultured for 16-20hrs and the presence of pronuclei determined under microscopy. Eggs having one or more pronuclei were given as activated.RESULTS: After in vitro culture, 139 out of 159 GV oocytes matured to the MII stage within 48hrs of culture (87.4%). Significantly more ova were mature at IVM2 than at IVM1 (P=0.039). Namely, at IVM1, eighty-nine ova matured from GV to MII stages (55.9%), whereas at IVM2, 50 out of 70 remaining oocytes completed maturation to MII (71.4%). On the other hand, following IVM period, eleven GV oocytes (6.9%) did not progress on oogenesis and 5.7% oocytes (n=9) just partially matured to the MI stage. In order to study the developmental competence of spontaneously in vitro matured oocytes, 31 MII ova were activated artificially. After overnight culture, 27 eggs had almost an identifiable pronucleus (87.1%). Comparable activation rates were obtained, regardless the maturation timing (IVM1: 16/19 and IVM2: 11/12; P=0.95). Twenty-two eggs (81.5%) had one single pronucleus.CONCLUSIONS: High proportion of human GV oocytes, recovered from ovarian hyperstimulated cycles were able to mature spontaneously up to the MII stage, within 48hrs of in vitro maturation in hTF medium. Our preliminary data did also suggest that most of in vitro matured MII oocytes were competent to reassume the cell cycle and to start a haploid embryo development. OBJECTIVE: After controlled ovarian hyperstimulation, most of recovered ova are mature at the metaphase II (MII: 85%) while others immature, being at either the metaphase I (MI: 4%) or the GV stage (11%). To study the nuclear and cytoplasmic competence of GV oocytes was aimed here. DESIGN: Experimental research, approved by the Ethical Comitee of the Instituto Universitario IVI, Law14/2003, Spain. MATERIALS AND METHODS: A total of 159 GV oocytes from stimulated cycles were donated to this research. Individual GV oocytes were matured in 50μL hTF medium for 2 days. Progression on nuclear maturation was assessed twice along the in vitro maturation (IVM) period; at IVM1 (20-24hrs) and at IVM2 (44-48hrs). At these check points, ova were checked for the presence of a nucleus (GV), absence of nucleus nor polar body (MI) or absence of nucleus, but presence of the 1st polar body in the perivitelline space (MII). To determine the cytoplasmic competence, 31 MII ova were activated artificially. Briefly, they were incubated serial in 5μM A23187 (5min) and 10μg/mL puromycin (6hrs). After rinsing, they were cultured for 16-20hrs and the presence of pronuclei determined under microscopy. Eggs having one or more pronuclei were given as activated. RESULTS: After in vitro culture, 139 out of 159 GV oocytes matured to the MII stage within 48hrs of culture (87.4%). Significantly more ova were mature at IVM2 than at IVM1 (P=0.039). Namely, at IVM1, eighty-nine ova matured from GV to MII stages (55.9%), whereas at IVM2, 50 out of 70 remaining oocytes completed maturation to MII (71.4%). On the other hand, following IVM period, eleven GV oocytes (6.9%) did not progress on oogenesis and 5.7% oocytes (n=9) just partially matured to the MI stage. In order to study the developmental competence of spontaneously in vitro matured oocytes, 31 MII ova were activated artificially. After overnight culture, 27 eggs had almost an identifiable pronucleus (87.1%). Comparable activation rates were obtained, regardless the maturation timing (IVM1: 16/19 and IVM2: 11/12; P=0.95). Twenty-two eggs (81.5%) had one single pronucleus. CONCLUSIONS: High proportion of human GV oocytes, recovered from ovarian hyperstimulated cycles were able to mature spontaneously up to the MII stage, within 48hrs of in vitro maturation in hTF medium. Our preliminary data did also suggest that most of in vitro matured MII oocytes were competent to reassume the cell cycle and to start a haploid embryo development." @default.
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• W2100084351 date "2008-09-01" @default.
• W2100084351 modified "2023-09-26" @default.
• W2100084351 title "Successful spontaneous in vitro maturation of human germinal vesicle (GV) oocytes, recovered from stimulated cycles" @default.
• W2100084351 doi "https://doi.org/10.1016/j.fertnstert.2008.07.1645" @default.
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