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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 st...

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Autores principales: Anchan, Raymond, Gerami-Naini, Behzad, Lindsey, Jennifer S., Ho, Joshua W. K., Kiezun, Adam, Lipskind, Shane, Ng, Nicholas, LiCausi, Joseph A., Kim, Chloe S., Brezina, Paul, Tuschl, Thomas, Maas, Richard, Kearns, William G., Williams, Zev
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4353623/
https://www.ncbi.nlm.nih.gov/pubmed/25751620
http://dx.doi.org/10.1371/journal.pone.0119275
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author Anchan, Raymond
Gerami-Naini, Behzad
Lindsey, Jennifer S.
Ho, Joshua W. K.
Kiezun, Adam
Lipskind, Shane
Ng, Nicholas
LiCausi, Joseph A.
Kim, Chloe S.
Brezina, Paul
Tuschl, Thomas
Maas, Richard
Kearns, William G.
Williams, Zev
author_facet Anchan, Raymond
Gerami-Naini, Behzad
Lindsey, Jennifer S.
Ho, Joshua W. K.
Kiezun, Adam
Lipskind, Shane
Ng, Nicholas
LiCausi, Joseph A.
Kim, Chloe S.
Brezina, Paul
Tuschl, Thomas
Maas, Richard
Kearns, William G.
Williams, Zev
author_sort Anchan, Raymond
collection PubMed
description 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.
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spelling pubmed-43536232015-03-17 Efficient Differentiation of Steroidogenic and Germ-Like Cells from Epigenetically-Related iPSCs Derived from Ovarian Granulosa Cells Anchan, Raymond Gerami-Naini, Behzad Lindsey, Jennifer S. Ho, Joshua W. K. Kiezun, Adam Lipskind, Shane Ng, Nicholas LiCausi, Joseph A. Kim, Chloe S. Brezina, Paul Tuschl, Thomas Maas, Richard Kearns, William G. Williams, Zev PLoS One Research Article 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. Public Library of Science 2015-03-09 /pmc/articles/PMC4353623/ /pubmed/25751620 http://dx.doi.org/10.1371/journal.pone.0119275 Text en © 2015 Anchan et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Anchan, Raymond
Gerami-Naini, Behzad
Lindsey, Jennifer S.
Ho, Joshua W. K.
Kiezun, Adam
Lipskind, Shane
Ng, Nicholas
LiCausi, Joseph A.
Kim, Chloe S.
Brezina, Paul
Tuschl, Thomas
Maas, Richard
Kearns, William G.
Williams, Zev
Efficient Differentiation of Steroidogenic and Germ-Like Cells from Epigenetically-Related iPSCs Derived from Ovarian Granulosa Cells
title Efficient Differentiation of Steroidogenic and Germ-Like Cells from Epigenetically-Related iPSCs Derived from Ovarian Granulosa Cells
title_full Efficient Differentiation of Steroidogenic and Germ-Like Cells from Epigenetically-Related iPSCs Derived from Ovarian Granulosa Cells
title_fullStr Efficient Differentiation of Steroidogenic and Germ-Like Cells from Epigenetically-Related iPSCs Derived from Ovarian Granulosa Cells
title_full_unstemmed Efficient Differentiation of Steroidogenic and Germ-Like Cells from Epigenetically-Related iPSCs Derived from Ovarian Granulosa Cells
title_short Efficient Differentiation of Steroidogenic and Germ-Like Cells from Epigenetically-Related iPSCs Derived from Ovarian Granulosa Cells
title_sort efficient differentiation of steroidogenic and germ-like cells from epigenetically-related ipscs derived from ovarian granulosa cells
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4353623/
https://www.ncbi.nlm.nih.gov/pubmed/25751620
http://dx.doi.org/10.1371/journal.pone.0119275
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