Efficient differentiation of steroidogenic and germ-like cells from epigenetically-related iPSCs derived from ovarian granulosa cells

PloS One
Raymond AnchanZev Williams

Abstract

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-E...Continue Reading

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Citations

Feb 14, 2016·Stem Cells and Development·Aurélie MoukaLucie Tosca
Oct 19, 2017·Current Protocols in Human Genetics·Sophia SalasRaymond M Anchan
Jun 14, 2019·Clinical Medicine Insights. Reproductive Health·Taichi AkahoriJonathan L Tilly
Nov 7, 2019·International Journal of Molecular Sciences·Daniel Rodríguez Gutiérrez, Anna Biason-Lauber
May 8, 2020·International Journal of Molecular Sciences·Erica SilvestrisAngelo Virgilio Paradiso
Jan 30, 2021·Stem Cells International·Maisumu GulimiheranmuJunmei Zhou

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Methods Mentioned

BETA
SMA
PCR

Software Mentioned

BeadStudio
Ingenuity Pathways Analysis ( IPA )
R
limma
lumi
Illumina

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