Pup birth from mouse oocytes in preantral follicles derived from vitrified and warmed ovaries followed by in vitro growth, in vitro maturation, and in vitro fertilization

ObjectiveTo determine an optimum condition for vitrification of various animal species and to examine the developing capacity into pups of mouse oocytes in preantral follicles originating from cryopreserved ovaries.DesignExperimental animal study.SettingLaboratory environment.Animal(s)Normal (C57BL/6 × DBA2) F1 mice.Intervention(s)Vitrification of animal ovaries using polyester sheets as a storage device; collection of oocyte–granulosa cell complexes by enzymatic treatment; and in vitro growth (IVG), in vitro maturation (IVM), and IVF, with embryo transplantation to pseudopregnant mice.Main Outcome Measure(s)Histological analysis of vitrified and warmed ovaries from several animal species; measurement of successful rates in IVG, IVM, and IVF of mouse oocytes in oocyte–granulosa cell complexes collected from the vitrified and warmed ovaries; and achievement of pup birth.Result(s)The vitrification method used was effective for storage of various animal ovaries. The oocytes enclosed in preantral follicles that were yielded from the vitrified–warmed ovaries preserved the capacity for developing into pups after IVG, IVM, and IVF.Conclusion(s)The findings indicated that ovarian cryopreservation by vitrification and subsequent IVG, IVM, and IVF are promising methods for restoring young cancer patients’ fertility. The practical information presented here is applicable to human ovarian tissues.