Efficient differentiation of steroidogenic and germ-like cells from epigenetically-related iPSCs derived from ovarian granulosa cells.
To explore restoration of ovarian function using epigenetically-related, induced pluripotent stem cells (iPSCs), we functionally evaluated the epigenetic memory of novel iPSC lines, derived from mouse and human ovarian granulosa cells (GCs) using c-Myc, Klf4, Sox2 and Oct4 retroviral vectors. The stem cell identity of the mouse and human GC-derived iPSCs (mGriPSCs, hGriPSCs) was verified by demonstrating embryonic stem cell (ESC) antigen expression using immunocytochemistry and RT-PCR analysis, as well as formation of embryoid bodies (EBs) and teratomas that are capable of differentiating into cells from all three germ layers. GriPSCs' gene expression profiles associate more closely with those of ESCs than of the originating GCs as demonstrated by genome-wide analysis of mRNA and microRNA. A comparative analysis of EBs generated from three different mouse cell lines (mGriPSCs; fibroblast-derived iPSC, mFiPSCs; G4 embryonic stem cells, G4 mESCs) revealed that differentiated mGriPSC-EBs synthesize 10-fold more estradiol (E2) than either differentiated FiPSC- or mESC-EBs under identical culture conditions. By contrast, mESC-EBs primarily synthesize progesterone (P4) and FiPSC-EBs produce neither E2 nor P4. Differentiated mGriPSC-EBs also express ovarian markers (AMHR, FSHR, Cyp19a1, ER and Inha) as well as markers of early gametogenesis (Mvh, Dazl, Gdf9, Boule and Zp1) more frequently than EBs of the other cell lines. These results provide evidence of preferential homotypic differentiation of mGriPSCs into ovarian cell types. Collectively, our data support the hypothesis that generating iPSCs from the desired tissue type may prove advantageous due to the iPSCs' epigenetic memory.
Division of Reproductive Endocrinology and Infertility, Department of Obstetrics, Gynecology and Reproductive Biology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.
Raymond
R
Anchan
Division of Genetics, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.
Behzad
B
Gerami-Naini
Division of Reproductive Endocrinology and Infertility, Department of Obstetrics, Gynecology and Reproductive Biology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.
Jennifer S
JS
Lindsey
Victor Chang Cardiac Research Institute and the University of New South Wales, Sydney, New South Wales, Australia.
Joshua W K
JW
Ho
Computational Methods Development, Cancer Genome Analysis, Broad Institute of MIT and Harvard, Cambridge, Massachusetts, United States of America.
Adam
A
Kiezun
Division of Reproductive Endocrinology and Infertility, Department of Obstetrics, Gynecology and Reproductive Biology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.
Shane
S
Lipskind
Division of Reproductive Endocrinology and Infertility, Department of Obstetrics, Gynecology and Reproductive Biology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.
Nicholas
N
Ng
Division of Reproductive Endocrinology and Infertility, Department of Obstetrics, Gynecology and Reproductive Biology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.
Joseph A
JA
LiCausi
Division of Genetics, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.
Chloe S
CS
Kim
Department of Gynecology and Obstetrics, John's Hopkins University School of Medicine, Baltimore, Maryland, United States of America, and Center for Preimplantation Genetics, LabCorp, Rockville, Maryland, United States of America.
Paul
P
Brezina
Laboratory of RNA Molecular Biology, Howard Hughes Medical Institute, The Rockefeller University, New York, New York, United States of America.
Thomas
T
Tuschl
Division of Genetics, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.
Richard
R
Maas
Department of Gynecology and Obstetrics, John's Hopkins University School of Medicine, Baltimore, Maryland, United States of America, and Center for Preimplantation Genetics, LabCorp, Rockville, Maryland, United States of America.
William G
WG
Kearns
Division of Reproductive Endocrinology and Infertility, Department of Obstetrics, and Gynecology and Women's Health, Albert Einstein College of Medicine, Bronx, New York, United States of America.
Zev
Z
Williams
D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D058732:Embryoid Bodies / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D044127:Epigenesis, Genetic; D004958:Estradiol / Q000378:metabolism*; D005260:Female; D005855:Germ Layers / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D006107:Granulosa Cells / Q000166:cytology*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism*; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000235:genetics; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D011374:Progesterone / Q000378:metabolism*; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics; D012190:Retroviridae / Q000235:genetics / Q000276:immunology; D055748:SOXB1 Transcription Factors / Q000235:genetics
C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; D050814:Octamer Transcription Factor-3; D016271:Proto-Oncogene Proteins c-myc; D055748:SOXB1 Transcription Factors; D011374:Progesterone; D004958:Estradiol