Journal
MOLECULAR THERAPY
Volume 20, Issue 2, Pages 408-416Publisher
CELL PRESS
DOI: 10.1038/mt.2011.258
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Funding
- NSF [MRI-DBI 0923559]
- Loma Linda University School of Medicine
- Loma Linda University Department of Medicine
- Loma Linda University GRASP
- DOD [W81XWH-11-1-0607]
- USAMRAA [W81XWH-08-1-0697]
- Division of Anatomy, the Department of Basic Sciences, the Center for Health Disparities and Molecular Medicine at Loma Linda University
- Radiation Research Laboratories in the Department of Radiation Medicine at Loma Linda University
- Div Of Biological Infrastructure
- Direct For Biological Sciences [923559] Funding Source: National Science Foundation
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The reprogramming of cord blood (CB) cells into induced pluripotent stem cells (iPSCs) has potential applications in regenerative medicine by converting CB banks into iPSC banks for allogeneic cell replacement therapy. Therefore, further investigation into novel approaches for efficient reprogramming is necessary. Here, we show that the lentiviral expression of OCT4 together with SOX2 (OS) driven by a strong spleen focus-forming virus (SFFV) promoter in a single vector can convert 2% of CB CD34(+) cells into iPSCs without additional reprogramming factors. Reprogramming efficiency was found to be critically dependent upon expression levels of OS. To generate transgene-free iPSCs, we developed an improved episomal vector with a woodchuck post-transcriptional regulatory element (Wpre) that increases transgene expression by 50%. With this vector, we successfully generated transgene-free iPSCs using OS alone. In conclusion, high-level expression of OS alone is sufficient for efficient reprogramming of CB CD34(+) cells into iPSCs. This report is the first to describe the generation of transgene-free iPSCs with the use of OCT4 and SOX2 alone. These findings have important implications for the clinical applications of iPSCs.
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