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Embryonic Stem Cells/Induced Pluripotent Stem Cells
Article first published online: 26 OCT 2010
Copyright © 2010 AlphaMed Press
Volume 28, Issue 10, pages 1728–1740, October 2010
How to Cite
Somers, A., Jean, J.-C., Sommer, C. A., Omari, A., Ford, C. C., Mills, J. A., Ying, L., Sommer, A. G., Jean, J. M., Smith, B. W., Lafyatis, R., Demierre, M.-F., Weiss, D. J., French, D. L., Gadue, P., Murphy, G. J., Mostoslavsky, G. and Kotton, D. N. (2010), Generation of Transgene-Free Lung Disease-Specific Human Induced Pluripotent Stem Cells Using a Single Excisable Lentiviral Stem Cell Cassette. STEM CELLS, 28: 1728–1740. doi: 10.1002/stem.495
Author contributions: A.S., J.C.J., C.A.S., C.C.F., J.A.M., and L.Y.: collection and assembly of data, data analysis and interpretation; A.O., A.G.S., and B.W.S.: collection and assembly of data; R.L., D.J.W., and M.F.D.: supply reagents; D.L.F. and P.G.: conception and design, data analysis and interpretation; G.J.M.: conception and design, collection and assembly of data, data analysis and interpretation, manuscript writing; G.M. and D.N.K.: conception and design, data analysis and interpretation, manuscript writing, final approval of manuscript. A.S., J.C.J., and C.A.S. contributed equally to this article.
Disclosure of potential conflicts of interest is found at the end of this article.
First published online in STEM CELLS EXPRESS August 16, 2010.
- Issue published online: 26 OCT 2010
- Article first published online: 26 OCT 2010
- Accepted manuscript online: 16 AUG 2010 12:00AM EST
- Manuscript Accepted: 1 AUG 2010
- Manuscript Received: 30 MAR 2010
- Human induced pluripotent stem cells;
- Human excisable single lentiviral vector;
- Stem cell cassette;
- Lung disease-specific iPSC;
- Cystic fibrosis;
- Alpha-1 antitrypsin deficiency;
- Sickle cell;
The development of methods to achieve efficient reprogramming of human cells while avoiding the permanent presence of reprogramming transgenes represents a critical step toward the use of induced pluripotent stem cells (iPSC) for clinical purposes, such as disease modeling or reconstituting therapies. Although 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 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 to accomplish efficient reprogramming of normal or diseased skin fibroblasts obtained from humans of virtually any age. Simultaneous transfer of either three or four reprogramming factors into human target cells using this single vector allows derivation of human iPSC containing a single excisable viral integration that on 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, α-1 antitrypsin deficiency-related emphysema, scleroderma, 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. STEM CELLS 2010;28:1728–1740