title: Allotransplantation of cryopreserved prepubertal mouse ovaries restored puberty and fertility without affecting methylation profile of Snrpn-DMR.
abstract: OBJECTIVE
  To evaluate the genetic safety of vitrification on the methylation imprints and the development and fertility potential of prepubertal mouse ovaries.
  
  
  METHODS
  Experimental animal study.
  
  
  METHODS
  University-based fertility center.
  
  
  METHODS
  Institute of Cancer Research (ICR) 10-day-old female mice, 10-week-old adult female mice, and 12-week-old adult male mice.
  
  
  METHODS
  Vitrification of juvenile mouse ovaries was performed using ED20 and EG5.5/30 solutions followed by retrieval of fresh and vitrified-warmed germinal vesicle (GV) oocytes for Snrpn differentially methylated regions (DMR) methylation analyses, collection of mature oocytes from superovulated ovarian grafts, in vitro fertility(IVF), and early embryonic development after heterotopic allotransplantation.
  
  
  METHODS
  Analysis of methylation status of Snrpn-DMR, percentage of fertilization, and blastocysts formation.
  
  
  RESULTS
  Methylation status of Snrpn-DMR from vitrified-warmed GV oocytes did not show significant alteration compared with that of controls, although a significant reduction of viable oocytes was observed. Puberty as well as endocrine function was restored, and no significant difference was shown in number of follicles, percentage of mice retaining fertility, and blastocyst formation among three groups.
  
  
  CONCLUSIONS
  Our study proved that vitrification of prepubertal mouse ovaries did not alter the methylation profile of Snrpn-DMR and subsequent allotransplantation; IVF could restore the development and fertility potential.
authors.affiliation: Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, National Research Center for Assisted Reproductive Technology and Reproductive Genetics, Key Laboratory for Reproductive Endocrinology of Ministry of Education, Shandong Provincial Key Laboratory of Reproductive Medicine, Jinan, People's Republic of China; Key Laboratory of Fertility Preservation and Maintenance of Ningxia Medical University and Ministry of Education of China, Yinchuan, People's Republic of China.
authors.forename: Hong-Yan
authors.initials: HY
authors.lastname: Wang
authors.affiliation: Key Laboratory of Fertility Preservation and Maintenance of Ningxia Medical University and Ministry of Education of China, Yinchuan, People's Republic of China.
authors.forename: Yun-Hong
authors.initials: YH
authors.lastname: Li
authors.affiliation: Research Center of Developmental Biology, College of Basic Medical Sciences, Second Military Medical University, Shanghai, People's Republic of China.
authors.forename: Lei
authors.initials: L
authors.lastname: Sun
authors.affiliation: Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, National Research Center for Assisted Reproductive Technology and Reproductive Genetics, Key Laboratory for Reproductive Endocrinology of Ministry of Education, Shandong Provincial Key Laboratory of Reproductive Medicine, Jinan, People's Republic of China.
authors.forename: Xuan
authors.initials: X
authors.lastname: Gao
authors.affiliation: Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, National Research Center for Assisted Reproductive Technology and Reproductive Genetics, Key Laboratory for Reproductive Endocrinology of Ministry of Education, Shandong Provincial Key Laboratory of Reproductive Medicine, Jinan, People's Republic of China.
authors.forename: Li
authors.initials: L
authors.lastname: You
authors.affiliation: Key Laboratory of Fertility Preservation and Maintenance of Ningxia Medical University and Ministry of Education of China, Yinchuan, People's Republic of China.
authors.forename: Yin
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, National Research Center for Assisted Reproductive Technology and Reproductive Genetics, Key Laboratory for Reproductive Endocrinology of Ministry of Education, Shandong Provincial Key Laboratory of Reproductive Medicine, Jinan, People's Republic of China.
authors.forename: Jing-Long
authors.initials: JL
authors.lastname: Ma
authors.affiliation: Center for Reproductive Medicine, Provincial Hospital Affiliated to Shandong University, National Research Center for Assisted Reproductive Technology and Reproductive Genetics, Key Laboratory for Reproductive Endocrinology of Ministry of Education, Shandong Provincial Key Laboratory of Reproductive Medicine, Jinan, People's Republic of China. Electronic address: chenzijiang@hotmail.com.
authors.forename: Zi-Jiang
authors.initials: ZJ
authors.lastname: Chen
mesh_terms: D000818:Animals; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology; D019175:DNA Methylation / Q000502:physiology*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D047108:Embryonic Development / Q000502:physiology; D019855:Ethylene Glycol / Q000494:pharmacology; D005260:Female; D005298:Fertility / Q000502:physiology*; D005307:Fertilization in Vitro / Q000379:methods; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D023421:Models, Animal; D010053:Ovary / Q000166:cytology / Q000502:physiology* / Q000637:transplantation*; D011627:Puberty / Q000502:physiology*; D012741:Sexual Maturation / Q000502:physiology; D013395:Sucrose / Q000494:pharmacology; D014184:Transplantation, Homologous; D058989:Vitrification; D055517:snRNP Core Proteins / Q000235:genetics*
chemical_list: D003451:Cryoprotective Agents; D055517:snRNP Core Proteins; D013395:Sucrose; D019855:Ethylene Glycol; D004121:Dimethyl Sulfoxide