Abstract
The generation of iPS cells gives an opportunity to use patient-specific somatic cells which are a valuable source for disease modeling and drug discovery. To promote these studies, it is important to make iPS cells from easily accessible and less invasive tissues like blood. Here, we describe the basic method to generate human iPS cells from adult peripheral blood. After the isolation of mononuclear cells, a combination of cytokines stimulates the expansion of hematopoietic stem/progenitor population, which is the main target of this protocol. The cells are transduced with plasmid mixture encoding reprogramming factors. In most cases, the plasmids are lost during the establishment of iPS clones.
Materials and Reagents
Equipment
Procedure
Day 0
Day 5
Day 6
Day 8, 10, and 12
Day 14-
Day 25 to 35
Notes
Recipes
Acknowledgments
We thank T. Aoi, K. Takahashi, M. Nakagawa and Y. Yoshida for scientific discussion; M. Narita, T. Ichisaka and M. Ohuchi for technical assistance; R. Kato, E. Nishikawa, S. Takeshima, and Y. Ohtsu for administrative assistance; and Drs. H. Niwa (RIKEN) and J. Miyazaki (Osaka University) for the CAG promoter. This study was supported in part by a grant from the Program for Promotion of Fundamental Studies in Health Sciences of National Institute of Biomedical Innovation, a grant from the Leading Project of Ministry of Education, Culture, Sports, Science and Technology (MEXT), a grant from Funding Program for World-Leading Innovative Research and Development on Science and Technology (FIRST Program) of Japan Society for the Promotion of Science, Grants-in-Aid for Scientific Research of Japan Society for the Promotion of Science and MEXT (to S.Y.), and Grants-in-Aid for Scientific Research for Young Scientists B (to K.O.).
References
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