Journal
STEM CELLS
Volume 31, Issue 3, Pages 458-466Publisher
WILEY
DOI: 10.1002/stem.1293
Keywords
Induced pluripotent stem cells; Reprogramming; Plasmid; Peripheral blood; T cells; Cord blood
Categories
Funding
- Program for Promotion of Fundamental Studies in Health Sciences of National Institute of Biomedical Innovation
- Ministry of Education, Culture, Sports, Science and Technology (MEXT)
- Funding Program for World-Leading Innovative Research and Development on Science and Technology (FIRST Program) of Japan Society for the Promotion of Science
- Japan Society for the Promotion of Science
- MEXT
- Grants-in-Aid for Scientific Research [24790276] Funding Source: KAKEN
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The generation of induced pluripotent stem cells (iPSCs) provides the opportunity to use patient-specific somatic cells, which are a valuable source for disease modeling and drug discovery. To promote research involving these cells, it is important to make iPSCs from easily accessible and less invasive tissues, like blood. We have recently reported the efficient generation of human iPSCs from adult fibroblasts using a combination of plasmids encoding OCT3/4, SOX2, KLF4, L-MYC, LIN28, and shRNA for TP53. We herein report a modified protocol enabling efficient iPSC induction from CD34(+) cord blood cells and from peripheral blood isolated from healthy donors using these plasmid vectors. The original plasmid mixture could induce iPSCs; however, the efficiency was low. The addition of EBNA1, an essential factor for episomal amplification of the vectors, by an extra plasmid greatly increased the efficiency of iPSC induction, especially when the induction was performed from alpha beta T cells. This improvement enabled the establishment of blood-derived iPSCs from seven healthy donors ranging in age from their 20s to their 60s. This induction method will be useful for the derivation of patient-specific integration-free iPSCs and would also be applicable to the generation of clinical-grade iPSCs in the future. STEM CELLS 2013;31:458-466
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