Quantification of Retinogenesis in 3D Cultures Reveals Epigenetic Memory and Higher Efficiency in iPSCs Derived from Rod Photoreceptors.
Quantification of Retinogenesis in 3D Cultures Reveals Epigenetic Memory and Higher Efficiency in iPSCs Derived from Rod Photoreceptors.
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DOI：
10.1016/j.stem.2015.05.015
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发表时间：
2015-07-02
期刊：
影响因子：
23.9
通讯作者：
Dyer MA
中科院分区：
文献类型：
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作者：
Hiler D;Chen X;Hazen J;Kupriyanov S;Carroll PA;Qu C;Xu B;Johnson D;Griffiths L;Frase S;Rodriguez AR;Martin G;Zhang J;Jeon J;Fan Y;Finkelstein D;Eisenman RN;Baldwin K;Dyer MA
治疗视网膜变性的细胞疗法目前正在临床试验中进行测试。然而，目前尚不清楚干细胞的来源对于产生适合移植的分化细胞是否重要。为了测试这一点，我们产生了iPSC鼠视杆细胞（r-iPSC），并使用称为STEM-RET的标准化定量方案对其制造视网膜的能力进行了评分。我们发现，r-iPSC在产生分化的视网膜方面比胚胎干细胞（ESC）或成纤维细胞衍生的iPSC（f-iPSC）更有效。来源于f-iPSC的视网膜具有无长突细胞和其他内核层细胞的减少。综合表观遗传学分析表明，DNA甲基化导致f-iPSC视网膜发生缺陷，并且视杆细胞特异性CTCF绝缘子蛋白结合位点可能促进r-iPSC中的视网膜原。总之，我们的数据表明，干细胞的来源对于在3D器官培养中产生视网膜神经元很重要。
Cell-based therapies to treat retinal degeneration are now being tested in clinical trials. However, it is not known if the source of stem cells is important for the production of differentiated cells suitable for transplantation. To test this, we generated iPSCs murine rod photoreceptors (r-iPSCs) and scored their ability to make retina using a standardized quantitative protocol called STEM-RET. We discovered that r-iPSCs were more efficient at producing differentiated retina than embryonic stem cells (ESCs) or fibroblast-derived iPSCs (f-iPSCs). Retinae derived from f-iPSCs had a reduction in amacrine cells and other inner nuclear layer cells. Integrated epigenetic analysis showed that DNA methylation contributes to the defects in f-iPSC retinogenesis and that rod specific CTCF insulator protein binding sites may promote retinogenes in r-iPSCs. Taken together, our data suggest that the source of stem cells are important for producing retinal neurons in 3D organ cultures.