Whole Ovary Vitrification for Fertility Preservation

The long term goal of this proposal is to develop a safeeffective and user friendly vitrification system for cryopreservation of whole human ovariesIn the U Sovergirls and women ageare diagnosed with cancer annuallyWhile improved diagnosis and treatment have led to increased survival ratesin premenopausal andin childhood cancer patientsdevastating side effects of cancer therapies include premature ovarian failureinfertility and menopause related health risksWhile cryopreservation of embryos and oocytes are effective and will remain as the most commonly used fertility preservation methodovarian cortex and whole ovary cryopreservation is the only option for future fertility in prepubertal patients and those who require immediate cancer treatmentCryopreservation of whole ovariescompared to ovarian tissuehas added advantages ofpreserving all classes of follicleswhich can be used for in vitro maturationlonger graft lifespanandfaster recovery supported by existing blood vessels after transplantation with vascular anastomosisHowevercurrent methods for whole ovary cryopreservation are suboptimal and there is no live birth in human from cryopreserved whole ovariesdespite it may be the best choice for prepubertal cancer patients for future fertility and intact endocrine function for decades without the help of exogenous hormonesThe proposed research is part ofst Century Medicine s more general program of developing superior cryopreservation systemincluding cryoprotective agentCPAsolutionsperfusion apparatushardware and softwarefor CPA loading and unloadingand liquid nitrogen storage unitsfor organsOur company has successfully demonstrated functional preservation of vitrified rabbit kidneys after transplantationThe pig and human ovaries are similar in size and smallergramscompared to rabbit kidneysgramswhich makes vitrification of pig and human ovaries more achievable than rabbit kidneysIn this SBIR Phase I studywe will establish optimal perfusion parameters and CPA combination that will allow the preservation of follicles and blood vessels in whole swine ovariesand design an ovary specific perfusion apparatusAimWe will also optimize cooling and warming ratesand determine whether radiofrequency heating can be utilized to improve efficiency and uniformity during rewarming of vitrified swine ovariesAimOvarian function will be evaluated in vitro by ovarian tissue culturehistology and immunohistochemistrypreantral follicle culturesurvivalantrum formationhormone productioncumulus oocyte complex cultureoocyte maturationResults from these studies will improve current technology in whole ovary cryopreservation and include a design and prototype of an automated vitrification systemFuture Phase II studies will focus on construction of the vitrification apparatus incorporating aspects of perfusioncoolingstorage and rewarming with a userfriendly computer interfaceThis novel system will be tested in vivoto demonstrate livebirth in the swine model after transplantationand in vitroto test ovarian function using human ovariesin Phase II One devastating side effect of cancer treatment is premature ovarian failure which can lead to infertility and long term health risks associated with menopauseCompared to existing fertility preservation methodsembryooocyte and ovarian tissue cryopreservationwhole ovary cryopreservation offers complete recovery of ovarian functionendocrine and fertilitywhich is important for youngespecially prepubertalcancer patientsHowevercurrentlyno live birth has been demonstrated from cryopreserved whole human ovaries due to the lack of perfusion system and optimal cryopreservation protocolsThe goals for this Phase I SBIR proposal are to establish optimal perfusion as well as cryopreservation conditions for the whole ovary and design a noveleffective and user friendly vitrification apparatususing the swine ovary as an animal model