High-efficiency RNA-based reprogramming of human primary fibroblasts.
High-efficiency RNA-based reprogramming of human primary fibroblasts.
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DOI：
10.1038/s41467-018-03190-3
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发表时间：
2018-02-21
影响因子：
16.6
通讯作者：
Bilousova G
中科院分区：
文献类型：
--
作者：
Kogut I;McCarthy SM;Pavlova M;Astling DP;Chen X;Jakimenko A;Jones KL;Getahun A;Cambier JC;Pasmooij AMG;Jonkman MF;Roop DR;Bilousova G
诱导性多能干细胞（iPSC）在再生医学方面有着巨大的前景;然而，其潜在的临床应用受到体细胞重编程效率低下的阻碍。在这里，我们表明，编码重编程因子的合成修饰mRNA和作为成熟miRNA模拟物递送的miRNA-367/302的协同活性极大地增强了人原代成纤维细胞向iPSC的重编程。这种协同活性取决于最佳的RNA转染方案和专门针对人原代成纤维细胞的培养条件。因此，我们现在可以从500个起始人原代新生儿成纤维细胞中产生多达4，019个iPSC集落，并重新编程高达90.7%的单个平板细胞，产生多个姐妹集落。这种方法在无饲养层条件下从各种人类患者的成纤维细胞中持续产生临床相关的无整合iPSC，并且可以适用于iPSC的临床翻译和研究重编程的生物学。诱导多能干细胞（iPSC）具有再生医学应用的潜力，但产生的效率非常低。在这里，作者显示了通过合成修饰的mRNA、成熟的miRNA模拟物和优化的培养方法的协同活性，将人原代成纤维细胞高效重编程为iPSC。
Induced pluripotent stem cells (iPSCs) hold great promise for regenerative medicine; however, their potential clinical application is hampered by the low efficiency of somatic cell reprogramming. Here, we show that the synergistic activity of synthetic modified mRNAs encoding reprogramming factors and miRNA-367/302s delivered as mature miRNA mimics greatly enhances the reprogramming of human primary fibroblasts into iPSCs. This synergistic activity is dependent upon an optimal RNA transfection regimen and culturing conditions tailored specifically to human primary fibroblasts. As a result, we can now generate up to 4,019 iPSC colonies from only 500 starting human primary neonatal fibroblasts and reprogram up to 90.7% of individually plated cells, producing multiple sister colonies. This methodology consistently generates clinically relevant, integration-free iPSCs from a variety of human patient’s fibroblasts under feeder-free conditions and can be applicable for the clinical translation of iPSCs and studying the biology of reprogramming. Induced pluripotent stem cells (iPSCs) have potential for regenerative medicine applications, but are generated with very low efficiency. Here, the authors show highly efficient reprogramming of human primary fibroblasts to iPSCs via the synergistic activity of synthetic modified mRNAs, mature miRNA mimics, and optimized culture methods.
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影响因子：
46.9
作者：
Schlaeger, Thorsten M.;Daheron, Laurence;Brickler, Thomas R.;Entwisle, Samuel;Chan, Karrie;Cianci, Amelia;DeVine, Alexander;Ettenger, Andrew;Fitzgerald, Kelly;Godfrey, Michelle;Gupta, Dipti;McPherson, Jade;Malwadkar, Prerana;Gupta, Manav;Bell, Blair;Doi, Akiko;Jung, Namyoung;Li, Xin;Lynes, Maureen S.;Brookes, Emily;Cherry, Anne B. C.;Demirbas, Didem;Tsankov, Alexander M.;Zon, Leonard I.;Rubin, Lee L.;Feinberg, Andrew P.;Meissner, Alexander;Cowan, Chad A.;Daley, George Q.
通讯作者：
Daley, George Q.
影响因子：
23.9
作者：
Buganim, Yosef;Markoulaki, Styliani;van Wietmarschen, Niek;Hoke, Heather;Wu, Tao;Ganz, Kibibi;Akhtar-Zaidi, Batool;He, Yupeng;Abraham, Brian J.;Porubsky, David;Kulenkampff, Elisabeth;Faddah, Dina A.;Shi, Linyu;Gao, Qing;Sarkar, Sovan;Cohen, Malkiel;Goldmann, Johanna;Nery, Joseph R.;Schultz, Matthew D.;Ecker, Joseph R.;Xiao, Andrew;Young, Richard A.;Lansdorp, Peter M.;Jaenisch, Rudolf
通讯作者：
Jaenisch, Rudolf
Notch抑制允许与癌基因无关的IPS细胞产生。
影响因子：
14.8
作者：
Ichida, Justin K.;Julia, T. C. W.;Williams, Luis A.;Carter, Ava C.;Shi, Yingxiao;Moura, Marcelo T.;Ziller, Michael;Singh, Sean;Amabile, Giovanni;Bock, Christoph;Umezawa, Akihiro;Rubin, Lee L.;Bradner, James E.;Akutsu, Hidenori;Meissner, Alexander;Eggan, Kevin
通讯作者：
Eggan, Kevin
影响因子：
4.6
作者：
Warren, Luigi;Ni, Yuhui;Wang, Jiwu;Guo, Xirong
通讯作者：
Guo, Xirong
影响因子：
--
作者：
van den Akker, Peter C.;Nijenhuis, Miranda;Pasmooij, Anna M. G.
通讯作者：
Pasmooij, Anna M. G.