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An Embryonic and Induced Pluripotent Stem Cell Model for Ovarian Granulosa Cell Development and Steroidogenesis

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Abstract

Embryoid bodies (EBs) can serve as a system for evaluating pluripotency, cellular differentiation, and tissue morphogenesis. In this study, we use EBs derived from mouse embryonic stem cells (mESCs) and human amniocyte-derived induced pluripotent stem cells (hAdiPSCs) as a model for ovarian granulosa cell (GC) development and steroidogenic cell commitment. We demonstrated that spontaneously differentiated murine EBs (mEBs) and human EBs (hEBs) displayed ovarian GC markers, such as aromatase (CYPI9AI), FOXL2, AMHR2, FSHR, and GJAI. Comparative microarray analysis identified both shared and unique gene expression between mEBs and the maturing mouse ovary. Gene sets related to gonadogenesis, lipid metabolism, and ovarian development were significantly overrepresented in EBs. Of the 29 genes, 15 that were differentially regulated in steroidogenic mEBs displayed temporal expression changes between embryonic, postnatal, and mature ovarian tissues by polymerase chain reaction. Importantly, both mEBs and hEBs were capable of gonadotropin-responsive estradiol (E2) synthesis in vitro (217-759 pg/mL). Live fluorescence-activated cell sorting-sorted AMHR2+ granulosa-like cells from mEBs continued to produce E2 after purification (15.3 pg/mL) and secreted significantly more E2 than AMHR2 cells (8.6 pg/mL, P <.05). We conclude that spontaneously differentiated EBs of both mESC and hAdiPSC origin can serve as a biologically relevant model for ovarian GC differentiation and steroidogenic cell commitment. These cells should be further investigated for therapeutic uses, such as stem cell-based hormone replacement therapy and in vitro maturation of oocytes.

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Author information

Author notes

  1. Behzad Gerami-Naini PhD

    Present address: Department of Diagnostic Sciences, School of Dental Medicine, Tufts University, Boston, MA, USA

  2. Jennifer L. Eaton

    Present address: Division of Reproductive Endocrinology and Fertility, Department of Obstetrics and Gynecology, Duke University School of Medicine, Durham, NC, USA

  3. Daniel O’Connell PhD

    Present address: Intellia Therapeutics, Inc, Cambridge, MA, USA

  4. Adam Kiezun PhD

    Present address: Amazon.com, Boston, MA, USA

Authors and Affiliations

  1. Division of Reproductive Endocrinology and Infertility, Department of Obstetrics, Gynecology and Reproductive Medicine, Brigham and Women’s Hospital and Harvard Medical School, 75 Francis Street, Boston, MA, 02115, USA

    Shane Lipskind MD, Jennifer S. Lindsey BS, Nicholas Ng BS, Parveen Parasar DVM, Michelle Ng PhD, Michael Nickerson BS & Raymond M. Anchan MD, PhD

  2. Division of Genetics, Department of Medicine, Brigham and Women’s Hospital and Harvard Medical School, Boston, MA, USA

    Behzad Gerami-Naini PhD, Jennifer L. Eaton, Daniel O’Connell PhD & Richard Maas MD, PhD

  3. Computational Methods Development, Cancer Genome Analysis, Broad Institute of MIT and Harvard, Cambridge, MA, USA

    Adam Kiezun PhD

  4. Victor Chang Cardiac Research Institute, University of New South Wales, Sydney, New South Wales, Australia

    Joshua W. K. Ho PhD

  5. Canary Center at Stanford for Early Cancer Detection, Stanford School of Medicine, Palo Alto, CA, USA

    Utkan Demirci PhD

  6. Affiliated Faculty, Harvard Stem Cell Institute, Cambridge, MA, USA

    Richard Maas MD, PhD & Raymond M. Anchan MD, PhD

Authors
  1. Shane Lipskind MD
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  2. Jennifer S. Lindsey BS
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  7. Joshua W. K. Ho PhD
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  14. Raymond M. Anchan MD, PhD
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Corresponding author

Correspondence to Raymond M. Anchan MD, PhD.

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Lipskind, S., Lindsey, J.S., Gerami-Naini, B. et al. An Embryonic and Induced Pluripotent Stem Cell Model for Ovarian Granulosa Cell Development and Steroidogenesis. Reprod. Sci. 25, 712–726 (2018). https://doi.org/10.1177/1933719117725814

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