Efficient Differentiation of Steroidogenic and Germ-Like Cells from Epigenetically-Related iPSCs Derived from Ovarian Granulosa Cells

被引：17

作者：

Anchan, Raymond ^{[1
]}

Gerami-Naini, Behzad ^{[2
]}

Lindsey, Jennifer S. ^{[1
]}

Ho, Joshua W. K. ^{[3
,4
]}

Kiezun, Adam ^{[5
]}

Lipskind, Shane ^{[1
]}

Ng, Nicholas ^{[1
]}

LiCausi, Joseph A. ^{[1
]}

Kim, Chloe S. ^{[2
]}

Brezina, Paul ^{[6
,7
]}

Tuschl, Thomas ^{[8
]}

Maas, Richard ^{[2
]}

Kearns, William G. ^{[6
,7
]}

Williams, Zev ^{[9
]}

机构：

[1] Harvard Univ, Brigham & Womens Hosp, Dept Obstet Gynecol & Reprod Biol, Div Reprod Endocrinol & Infertil,Med Sch, Boston, MA 02115 USA

[2] Harvard Univ, Brigham & Womens Hosp, Dept Med, Div Genet,Med Sch, Boston, MA 02115 USA

[3] Victor Chang Cardiac Res Inst, Sydney, NSW, Australia

[4] Univ New S Wales, Sydney, NSW, Australia

[5] Broad Inst MIT & Harvard, Computat Methods Dev, Canc Genome Anal, Cambridge, MA USA

[6] Johns Hopkins Univ, Sch Med, Dept Gynecol & Obstet, Baltimore, MD 21205 USA

[7] LabCorp, Ctr Preimplantat Genet, Rockville, MD USA

[8] Rockefeller Univ, Howard Hughes Med Inst, Lab RNA Mol Biol, New York, NY 10021 USA

[9] Albert Einstein Coll Med, Div Reprod Endocrinol & Infertil, Dept Obstet & Gynecol & Womens Hlth, Bronx, NY 10467 USA

来源：

PLOS ONE | 2015年 / 10卷 / 03期

关键词：

PLURIPOTENT STEM-CELLS; GENE-EXPRESSION; COPY NUMBER; TRANSDIFFERENTIATION; GENERATION; MEMORY; CHEMOTHERAPY; FIBROBLASTS; NUCLEAR; CUMULUS;

D O I：

10.1371/journal.pone.0119275

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

070301 [无机化学]; 070403 [天体物理学]; 070507 [自然资源与国土空间规划学]; 090105 [作物生产系统与生态工程];

摘要：

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.

引用

页数：21

共 60 条

[1]

Amniocytes can serve a dual function as a source of iPS cells and feeder layers [J].