Generation of induced pluripotent stem cells from human foetal fibroblasts using the Sleeping Beauty transposon gene delivery system

被引:38
作者
Davis, Richard P. [1 ,2 ]
Nemes, Csilla [4 ]
Varga, Eszter [3 ,4 ]
Freund, Christian [1 ]
Kosmidis, Georgios [1 ]
Gkatzis, Konstantinos [1 ]
de Jong, Danielle [5 ]
Szuhai, Karoly [5 ]
Dinnyes, Andras [3 ,4 ]
Mummery, Christine L. [1 ,2 ]
机构
[1] Leiden Univ, Med Ctr, Dept Anat & Embryol, NL-2333 ZC Leiden, Netherlands
[2] Netherlands Prote Inst, Utrecht, Netherlands
[3] Szent Istvan Univ, Mol Anim Biotechnol Lab, Godollo, Hungary
[4] BioTalentum Ltd, Godollo, Hungary
[5] Leiden Univ, Med Ctr, Dept Mol Cell Biol, NL-2333 ZC Leiden, Netherlands
关键词
Human Induced pluripotent stem cell; Sleeping Beauty transposon; Differentiation; Transfection methods; Nuclear reprogramming; HUMAN GENOME; DIFFERENTIATION; INDUCTION; MOUSE; VECTOR; MUTAGENESIS; INTEGRATION; INSERTION; EXCISION; DISEASE;
D O I
10.1016/j.diff.2013.06.002
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Transposon gene delivery systems offer an alternative, non-viral-based approach to generate induced pluripotent stem cells (iPSCs). Here we used the Sleeping Beauty (SB) transposon to generate four human iPSC lines from foetal fibroblasts. In contrast to other gene delivery systems, the SB transposon does not exhibit an integration bias towards particular genetic elements, thereby reducing the risk of insertional mutagenesis. Furthermore, unlike the alternative transposon piggyBac, SB has no SB-like elements within the human genome, minimising the possibility of mobilising endogenous transposon elements. All iPSC lines exhibited the expected characteristics of pluripotent human cells, including the ability to differentiate to derivatives of all three germ layers in vitro. Re-expression of the SB transposase in the iPSCs after reprogramming resulted in the mobilisation of some of the transposons. These results indicate that the SB transposon system is a useful addition to methods for generating human iPSCs, both for basic and applied biomedical research, and in the context of future therapeutic application. (C) 2013 International Society of Differentiation. Published by Elsevier B.V. All rights reserved.
引用
收藏
页码:30 / 37
页数:8
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