Efficient differentiation of steroidogenic and germ-like cells from epigenetically-related iPSCs derived from ovarian granulosa cells.
Efficient differentiation of steroidogenic and germ-like cells from epigenetically-related iPSCs derived from ovarian granulosa cells.
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DOI：
10.1371/journal.pone.0119275
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Williams Z
中科院分区：
文献类型：
--
作者：
Anchan R;Gerami-Naini B;Lindsey JS;Ho JW;Kiezun A;Lipskind S;Ng N;LiCausi JA;Kim CS;Brezina P;Tuschl T;Maas R;Kearns WG;Williams Z
为了探索使用表观遗传相关的诱导多能干细胞（iPSC）恢复卵巢功能，我们功能性地评估了使用c-Myc、Klf 4、Sox 2和Oct 4逆转录病毒载体从小鼠和人卵巢颗粒细胞（GC）衍生的新型iPSC系的表观遗传记忆。通过使用免疫细胞化学和RT-PCR分析证明胚胎干细胞（ESC）抗原表达以及能够分化成来自所有三个胚层的细胞的胚状体（EB）和畸胎瘤的形成来验证小鼠和人GC衍生的iPSC（mGriPSC、hGriPSC）的干细胞身份。GriPSC的基因表达谱与ESC的基因表达谱比原始GC的基因表达谱更密切相关，如通过mRNA和microRNA的全基因组分析所证明的。由三种不同小鼠细胞系（mGriPSC;成纤维细胞衍生的iPSC，mFiPSC; G4胚胎干细胞，G4 mESC）产生的EB的比较分析揭示，在相同培养条件下，分化的mGriPSC-EB合成的雌二醇（E2）比分化的FiPSC-或mESC-EB多10倍。相比之下，mESC-EB主要合成孕酮（P4），FiPSC-EB既不产生E2也不产生P4。分化的mGriPSC-EB也比其他细胞系的EB更频繁地表达卵巢标志物（AMHR、FSHR、Cyp 19 a1、ER和Inha）以及早期配子发生标志物（Mvh、Dazl、Gdf 9、Boule和Zp 1）。这些结果提供了mGriPSC优先同型分化为卵巢细胞类型的证据。总的来说，我们的数据支持这样的假设，即由于iPSC的表观遗传记忆，从所需的组织类型产生iPSC可能是有利的。
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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影响因子：
3.7
作者：
Brown M;Suryawanshi H;Hafner M;Farazi TA;Tuschl T
通讯作者：
Tuschl T
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Hafner, Markus;Renwick, Neil;Farazi, Thalia A.;Mihailovic, Aleksandra;Pena, John T. G.;Tuschl, Thomas
通讯作者：
Tuschl, Thomas
影响因子：
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