Parthenogenic Blastocysts Derived from Cumulus-Free In Vitro Matured Human Oocytes 英文参考文献.docVIP

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Parthenogenic Blastocysts Derived from Cumulus-Free In Vitro Matured Human Oocytes 英文参考文献.doc

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Parthenogenic Blastocysts Derived from Cumulus-Free In Vitro Matured Human Oocytes 英文参考文献

ParthenogenicBlastocystsDerivedfromCumulus-FreeIn VitroMaturedHumanOocytes SohyunL.McElroy1,2,JamesA.Byrne1,2,ShawnL.Chavez1,2,BarryBehr2,3,AaronJ.Hsueh2,LynnM. Westphal2,3,ReneeA.ReijoPera1,2 * 1CenterforHumanEmbryonicStemCellResearchandEducation,InstituteforStemCellBiologyandRegenerativeMedicine,StanfordUniversity,PaloAlto,California, United States of America, 2Department of Obstetrics and Gynecology, Stanford University, Palo Alto, California, United States of America, 3Division ofReproductive EndocrinologyandInfertility,StanfordHospitalandClinics,PaloAlto,California,UnitedStatesofAmerica Abstract Background: Approximately 20% of oocytes are classified as immature and discarded following intracytoplasmic sperm injection(ICSI)procedures.Theseoocytesareobtainedfromgonadotropin-stimulatedpatients,andareroutinelyremoved fromthecumuluscellswhichnormallywouldmaturetheoocytes.Giventhereadyaccesstothesehumanoocytes,they representapotentialresourceforbothclinicalandbasicscienceapplication.Howevercultureconditionsforthematuration ofcumulus-freeoocyteshavenotbeenoptimized.Weaimedtoimprovematurationconditionsforcumulus-freeoocytesvia culturewithovarianparacrine/autocrinefactorsidentifiedbysinglecellanalysis. Methodology/Principal Finding: Immature human oocytes were matured in vitro via supplementation with ovarian paracrine/autocrinefactorsthatwereselectedbasedonexpressionofligandsinthecumuluscellsandtheircorresponding receptors in oocytes. Matured oocytes were artificially activated to assess developmental competence. Gene expression profiles of parthenotes were compared to IVF/ICSI embryos at morula and blastocyst stages. Following incubation in mediumsupplementedwithovarianfactors(BDNF,IGF-I,estradiol,GDNF,FGF2andleptin),agreaterpercentageofoocytes demonstratednuclearmaturationandsubsequently,underwentparthenogenesisrelativetocontrol.Similarly,cytoplasmic maturationwasalsoimprovedasindicatedbydevelopmenttoblastocyststage.ParthenogenicblastocystsexhibitedmRNA expression