Generation of transgene-free lung disease-specific human induced pluripotent stem cells using a single excisable lentiviral stem cell cassette.
Generation of transgene-free lung disease-specific human induced pluripotent stem cells using a single excisable lentiviral stem cell cassette.
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DOI：
10.1002/stem.495
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发表时间：
2010-10
期刊：
影响因子：
5.2
通讯作者：
Kotton, Darrell N.
中科院分区：
文献类型：
--
作者：
Somers, Aba;Jean, Jyh-Chang;Sommer, Cesar A.;Omari, Amel;Ford, Christopher C.;Mills, Jason A.;Ying, Lei;Sommer, Andreia Gianotti;Jean, Jenny M.;Smith, Brenden W.;Lafyatis, Robert;Demierre, Marie-France;Weiss, Daniel J.;French, Deborah L.;Gadue, Paul;Murphy, George J.;Mostoslavsky, Gustavo;Kotton, Darrell N.
关键词：
开发实现人类细胞有效重编程同时避免重编程转基因永久存在的方法，代表着将诱导多能干细胞（iPSC）用于临床目的（例如疾病建模或重建疗法）的关键一步。虽然存在多种从小鼠细胞或新生儿正常人体组织中产生不含重编程转基因的 iPSC 的方法，但仍然需要足够有效的重编程系统，以实现从患有遗传性或退行性疾病的人类中广泛衍生疾病特异性 iPSC。在这里，我们报告了使用单一慢病毒“干细胞盒”载体的人源化版本，以实现对从几乎任何年龄的人类中获得的正常或患病皮肤成纤维细胞的有效重编程。使用该单一载体将 3 或 4 个重编程因子同时转移到人类靶细胞中，可以衍生出包含单个可切除病毒整合的人类 iPSC，在去除病毒整合后产生不含整合转基因的人类 iPSC。作为原理证明，我们在这里应用此策略从患有影响肺部上皮、内皮或间质室的各种疾病的个体中产生> 100个肺部疾病特异性 iPSC 系，这些疾病包括囊性纤维化、α-1抗胰蛋白酶缺乏相关的肺气肿、硬皮病 (SSc) 和镰状细胞病。此外，我们证明用这种方法产生的人 iPSC 能够在体外稳健分化为定形内胚层，即肺上皮的发育前体组织。
The development of methods to achieve efficient reprogramming of human cells while avoiding the permanent presence of reprogramming transgenes represents a critical step towards the use of induced pluripotent stem cells (iPSC) for clinical purposes, such as disease modeling or reconstituting therapies. While several methods exist for generating iPSC free of reprogramming transgenes from mouse cells or neonatal normal human tissues, a sufficiently efficient reprogramming system is still needed in order to achieve the widespread derivation of disease-specific iPSC from humans with inherited or degenerative diseases. Here we report the use of a humanized version of a single lentiviral ‘stem cell cassette’ vector in order to accomplish efficient reprogramming of normal or diseased skin fibroblasts obtained from humans of virtually any age. Simultaneous transfer of either 3 or 4 reprogramming factors into human target cells using this single vector allows derivation of human iPSC containing a single excisable viral integration, that upon removal generates human iPSC free of integrated transgenes. As a proof of principle, here we apply this strategy to generate >100 lung disease-specific iPSC lines from individuals with a variety of diseases affecting the epithelial, endothelial, or interstitial compartments of the lung, including cystic fibrosis, alpha-1 antitrypsin deficiency-related emphysema, scleroderma (SSc), and sickle cell disease. Moreover, we demonstrate that human iPSC generated with this approach have the ability to robustly differentiate into definitive endoderm in vitro, the developmental precursor tissue of lung epithelia.
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DOI：
10.1126/science.1162494
发表时间：
2008-11-07
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Stadtfeld M;Nagaya M;Utikal J;Weir G;Hochedlinger K
通讯作者：
Hochedlinger K
影响因子：
23.9
作者：
Heng, Jian-Chien Dominic;Feng, Bo;Ng, Huck-Hui
通讯作者：
Ng, Huck-Hui
影响因子：
64.8
作者：
Park, In-Hyun;Zhao, Rui;Daley, George Q.
通讯作者：
Daley, George Q.
影响因子：
64.5
作者：
Soldner F;Hockemeyer D;Beard C;Gao Q;Bell GW;Cook EG;Hargus G;Blak A;Cooper O;Mitalipova M;Isacson O;Jaenisch R
通讯作者：
Jaenisch R
影响因子：
46.9
作者：
Gouon-Evans, Valerie;Boussemart, Lise;Keller, Gordon
通讯作者：
Keller, Gordon