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Vitrification of In vitro-matured Oocytes:Effects of meiotic spindle Morphology on Clinical Outcome

Reproductive and Developmental Medicine 2020年第1期4卷 18-24页

作者： Rui-Huan Gu Zhi-Chao Li Jing-Wen Lang Hua Chen Yun Feng Song Guo Jing Fu Xiao-Xi Sun Yi-Juan Sun Shanghai Ji Ai Genetics and IVF Institute Obstetrics and Gynecology HospitalFudan UniversityShanghai 200011China Center for Reproductive Medicine Renji HospitalShanghai Jiao Tong University School of MedicineShanghai 200120China Reproductive Medical Center Ruijin HospitalShanghai Jiao Tong University School of MedicineShanghai 200025China Department of Gynaecology and Obstetrics Qianfoshan HospitalShandong UniversityJinan 250014China Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases Shanghai 200011China

Objective:The meiotic spindle controls chromosome movement and mediates various functions essential for fertilization and early postfertilization *** study aimed to examine whether vitrification causes meiotic damage in vitro-matured metaphase II(MII)human oocytes,and whether the meiotic spindle morphology influences the subsequent developmental ***:The spindle characteristics of MII human oocytes in vitro matured were studied before and after vitrification using PolScope imaging and immunofluorescence *** developmental competence of oocytes was also ***:A total of 419 human MII oocytes were obtained from 593 intracytoplasmic sperm injection cycles at our *** these oocytes,54 were used for immunofluorescence staining,whereas the other oocytes were examined by PolScope imaging and classified into three groups according to the meiotic spindle morphology:(A)normal morphology,(B)weak refraction and short meiotic spindle,and(C)no detectable meiotic *** three groups demonstrated statistically significant differences in terms of survival after ***,differences were not found in terms of oocyte chromosome structure and meiotic spindle morphology on immunofluorescence staining performed before and after *** survival,fertilization,and early embryonic development rates were significantly higher in Group A than in Groups B and C with or without *** vitrification had no effect on these metrics in Group A,Groups B and C demonstrated significantly lower fertilization and cleavage rates after vitrification/***:Screening for normal meiotic spindle morphology and chromosome configuration before vitrification may increase the yield of healthy viable oocytes for various assisted reproductive technologies.

关键词： Immunofluorescence Staining meiotic spindle Oocytes PolScope Imaging Vitrification

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Vitrification of-matured Oocytes: Effects of meiotic spindle Morphology on Clinical Outcome

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生殖与发育医学(英文) 2020年第1期 18-19-20-21-22-23-24页

Objective: The meiotic spindle controls chromosome movement and mediates various functions essential for fertilization and early postfertilization events. This study aimed to examine whether vitrification causes meiotic damagein vitro-matured metaphase II (MII) human oocytes, and whether the meiotic spindle morphology influences the subsequent developmental ***: The spindle characteristics of MII human oocytesin vitro matured were studied before and after vitrification using PolScope imaging and immunofluorescence staining. The developmental competence of oocytes was also ***: A total of 419 human MII oocytes were obtained from 593 intracytoplasmic sperm injection cycles at our hospital. Of these oocytes, 54 were used for immunofluorescence staining, whereas the other oocytes were examined by PolScope imaging and classified into three groups according to the meiotic spindle morphology: (A) normal morphology, (B) weak refraction and short meiotic spindle, and (C) no detectable meiotic spindle. The three groups demonstrated statistically significant differences in terms of survival after vitrification. However, differences were not found in terms of oocyte chromosome structure and meiotic spindle morphology on immunofluorescence staining performed before and after vitrification. Oocyte survival, fertilization, and early embryonic development rates were significantly higher in Group A than in Groups B and C with or without vitrification. While vitrification had no effect on these metrics in Group A, Groups B and C demonstrated significantly lower fertilization and cleavage rates after vitrification/***: Screening for normal meiotic spindle morphology and chromosome configuration before vitrification may increase the yield of healthy viable oocytes for various assisted reproductive technologies.

关键词： Immunofluorescence Staining meiotic spindle Oocytes PolScope Imaging Vitrification

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The Role of Brain-derived Neurotrophic Factor in Mouse Oocyte Maturation in vitro

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2010年第6期30卷 781-785页

作者：张玲李洁苏萍熊承良 Family Planning Research Institute and Center of Reproductive Medicine Tongji Medical CollegeHuazhong University of Science and Technology

Brain-derived neurotrophic factor (BDNF) can promote developmental competence in mammalian oocytes during in vitro maturation (IVM),but the role of BDNF in oocyte maturation at cellular level is not still *** this study,mouse cumulus-enclosed oocytes subjected to IVM were fertilized and cultured to blastocyst *** spindle configuration and cortical granules distribution during oocyte maturation in vitro were assessed by using immunofluorescence and laser confocal *** results showed that BDNF contributed to the complete preimplantation development of mouse oocytes compared to the control oocytes (13.78% vs.5.92%;Pmeiotic spindle areas were significantly smaller and the number of cytoplasmic microtubule organizing centers was greater than that in the control,and the percentages of oocytes showed spindles positioned near the oolemma and a well-formed cortical granule-free domain were significantly higher than that of the *** morphological characteristics of the BDNF-treated oocytes were much closer to the oocytes matured in vivo than those of the control *** conclusion,BDNF can promote the developmental competence of mouse IVM oocytes,by improving the meiotic spindle configuration and location and cortical granules distribution at meiosis Ⅰ.

关键词： brain-derived neurotrophic factor oocyte in vitro maturation meiotic spindle cortical granule

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Vitrification versus Slow Freezing of Human Oocytes: Effects on Ultrastructure and Developmental Potential

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Reproductive and Developmental Medicine 2018年第3期2卷 129-136页

作者： You-Zhu Li Na Li Xiao-Hong Yan Wei-Dong Zhou Yu-Lai Zhou Qiong-Hua Chen Rong-Feng Wu Center for Reproductive Medicine The First Affiliated Hospital of Xiamen UniversityXiamen 361003China Intensive Care Unit Xiamen Hongai HospitalXiamen 361003China College of Pharmacy Jilin UniversityChangchun 130021China

Objective:This study compared spindles,cytoskeleton,and developmental potential between vitrified and slow-frozen oocytes using PolScope and electron ***:Oocytes were randomly divided into control,slow freeze-thaw,and vitrification freeze-thaw groups(0,1,and 3 h).PolScope was used to observe spindles,angle of spindles to the first polar bodies,surface areas of oocytes,and lining and outer ret of zona *** and ultrastructure of oocytes were observed by scanning electron microscopy and transmission electron *** measures were used to characterize the impacts of two freezing methods on the developmental capacity of human ***:The visible frequency of spindle formation was 92.4%,56.4%,11.2%,24.8%,and 61.1%in control group,slow freeze-thaw group,and the three vitrification freeze-thaw groups(0,1,and 3 h),*** to oocytes in the slow freeze group,the angle of the spindle to the first polar body in oocytes in the 3-h vitrification freeze-thaw group was less(37.3°vs.68°,P=0.023).No significant differences were observed between the surface area of oocytes,or lining and outer ret of oocyte zona pellucida between freeze-thaw in these same two *** appeared ***,protrusions on oocyte surfaces were increased,and microvilli were laid down on the membrane surface in the 3-h vitrification freeze-thaw group in comparison to the slow-freeze *** comparisons showed better recovery of perivitelline space and mitochondria between the 3-h vitrified and slow-frozen ***(2PN)fertilization rate observed in the slow-freeze group(65.7%)was lower than the rate seen in controls(79.2%,P=0.041).No significant differences were observed in 2PN fertilization,cleavage,and blastocyst formation rates between the 3-h vitrification freeze-thaw and control ***:Results suggest that vitrification freeze-thaw for oocyte cryopreservation was a better choice than slow freeze-thaw.

关键词： Human Oocyte meiotic spindle Slow Freezing Vitrification Zona Pellucida

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