title: Embryonic stem cell-derived granulosa cells participate in ovarian follicle formation in vitro and in vivo.
abstract: Differentiating embryonic stem cells (ESCs) can form ovarian follicle-like structures in vitro, consisting of an oocyte-like cell surrounded by somatic cells capable of steroidogenesis. Using a dual-fluorescence reporter system in which mouse ESCs express green fluorescent protein (GFP) under the control of a germ cell-specific Pou5f1 gene promoter and red fluorescent protein (Discosoma sp red [DsRed]) driven by the granulosa cell-specific Forkhead box L2 (Foxl2) gene promoter, we first confirmed in vitro formation of follicle-like structures containing GFP-positive cells surrounded by DsRed-positive cells. Isolated DsRed-positive cells specified from ECSs exhibited a gene expression profile consistent with granulosa cells, as revealed by the detection of messenger RNAs (mRNAs) for Foxl2, follistatin (Fst), anti-Müllerian hormone (Amh), and follicle-stimulating hormone receptor (Fshr) as well as by production of both progesterone and estradiol. In addition, treatment of isolated DsRed-expressing cells with follicle-stimulating hormone (FSH) significantly increased estradiol production over basal levels, confirming the presence of functional FSH receptors in these cells. Last, ESC-derived DsRed-positive cells injected into neonatal mouse ovaries became incorporated within the granulosa cell layer of immature follicles. These studies demonstrate that Foxl2-expressing ovarian somatic cells derived in vitro from differentiating ESCs express granulosa cell markers, actively associate with germ cells in vitro, synthesize steroids, respond to FSH, and participate in folliculogenesis in vivo.
authors.affiliation: Vincent Center for Reproductive Biology, MGH Vincent Department of Obstetrics and Gynecology, Massachusetts General Hospital, Boston, MA, USA.
authors.forename: Dori C
authors.initials: DC
authors.lastname: Woods
authors.forename: Yvonne A R
authors.initials: YA
authors.lastname: White
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Niikura
authors.forename: Sorapop
authors.initials: S
authors.lastname: Kiatpongsan
authors.forename: Ho-Joon
authors.initials: HJ
authors.lastname: Lee
authors.forename: Jonathan L
authors.initials: JL
authors.lastname: Tilly
mesh_terms: D000818:Animals; D054304:Anti-Mullerian Hormone / Q000378:metabolism; D002454:Cell Differentiation; D018920:Coculture Techniques; D053595:Embryonic Stem Cells / Q000378:metabolism / Q000502:physiology*; D004958:Estradiol / Q000378:metabolism; D061252:Feeder Cells; D005260:Female; D005640:Follicle Stimulating Hormone / Q000378:metabolism; D038681:Follistatin / Q000235:genetics; D000074842:Forkhead Box Protein L2; D051858:Forkhead Transcription Factors / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D017930:Genes, Reporter; D006107:Granulosa Cells / Q000378:metabolism / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D006080:Ovarian Follicle / Q000378:metabolism / Q000502:physiology* / Q000637:transplantation; D010052:Ovariectomy; D010053:Ovary / Q000378:metabolism / Q000502:physiology* / Q000637:transplantation; D011374:Progesterone / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D013997:Time Factors; D014162:Transfection
chemical_list: D038681:Follistatin; D000074842:Forkhead Box Protein L2; D051858:Forkhead Transcription Factors; C498893:Foxl2 protein, mouse; D008164:Luminescent Proteins; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D012333:RNA, Messenger; C413663:fluorescent protein 583; D049452:Green Fluorescent Proteins; D011374:Progesterone; D004958:Estradiol; D054304:Anti-Mullerian Hormone; D005640:Follicle Stimulating Hormone
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