Article

A chemical platform for improved induction of human iPSCs.

Department of Chemistry, The Scripps Research Institute, La Jolla, California, USA.
Nature Methods (Impact Factor: 25.95). 11/2009; 6(11):805-8. DOI: 10.1038/nmeth.1393
Source: PubMed

ABSTRACT The slow kinetics and low efficiency of reprogramming methods to generate human induced pluripotent stem cells (iPSCs) impose major limitations on their utility in biomedical applications. Here we describe a chemical approach that dramatically improves (200-fold) the efficiency of iPSC generation from human fibroblasts, within seven days of treatment. This will provide a basis for developing safer, more efficient, nonviral methods for reprogramming human somatic cells.

Download full-text

Full-text

Available from: Simon Hilcove, Jun 29, 2015
3 Followers
 · 
241 Views
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Embryonic stem cells and induced pluripotent stem (iPS) cells are usually maintained on feeder cells derived from mouse embryonic fibroblasts (MEFs). In recent years, the cell culture of iPS cells under serum- and feeder-free conditions is gaining attention in overcoming the biosafety issues for clinical applications. In this study, we report on the use of multiple small-molecular inhibitors (i.e., CHIR99021, PD0325901, and Thiazovivin) to efficiently cultivate mouse iPS cells without feeder cells in a chemically-defined and serum-free condition. In this condition, we showed that mouse iPS cells are expressing the Nanog, Oct3/4, and SSEA-1 pluripotent markers, indicating that the culture condition is optimized to maintain the pluripotent status of iPS cells. Without these small-molecular inhibitors, mouse iPS cells required the adaptation period to start the stable cell proliferation. The application of these inhibitors enabled us the shortcut culture method for the cellular adaptation. This study will be useful to efficiently establish mouse iPS cell lines without MEF-derived feeder cells.
    Cytotechnology 03/2014; 67(2). DOI:10.1007/s10616-013-9686-8 · 1.45 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: A better understanding of the cellular and molecular mechanisms involved in the reprogramming of somatic cells is essential for further improvement of induced pluripotent stem (iPS) cell technology. In this study, we enriched for cells actively undergoing reprogramming at different time points by sorting the cells stained with a stem cell-selective fluorescent chemical probe CDy1 for their global gene expression analysis. Day-to-day comparison of differentially expressed genes showed highly dynamic and transient gene expressions during reprogramming, which were largely distinct from those of fully-reprogrammed cells. An unbiased analysis of functional regulation indicated robust modulation of concurrent programs at critical junctures. Globally, transcriptional programs involved in cell proliferation, morphology and signal transduction were instantly triggered as early as 3days-post-infection to prepare the cell for reprogramming but became somewhat muted in the final iPS cells. On the other hand, the highly coordinated metabolic reprogramming process was more gradually activated. Subsequent network analysis of differentially expressed genes indicated PDGF-BB as a core player in reprogramming which was verified by our gain- and loss-of-function experiments. As such, our study has revealed previously-unknown insights into the mechanisms of cellular reprogramming.
    Stem Cell Research 03/2014; 12(3):730-741. DOI:10.1016/j.scr.2014.03.002 · 3.91 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Human neurodegenerative disorders such as Alzheimer's disease and Parkinson's disease represent unmet medical need. There is no effective cure available on the market. Several novel therapeutic approaches targeting fundamental features of these disorders have been proposed during the last two decades. Cell therapy represents one of the most promising therapeutic avenues targeting different pathological traits of these disorders. However, there are some caveats that should be taken into the consideration including ethical issues and limited utilization for routine clinical practice. It is unlikely that cell therapy constitutes the 'magic bullet' therapeutic approach that would meet all therapeutic needs. However, in the future it can potentially bolster the effect of disease modifying drugs by improving the brain environment and regulation of inflammatory and neurotrophic pathways.
    Journal of Alzheimer's disease: JAD 08/2013; 37(2). DOI:10.3233/JAD-130572 · 3.61 Impact Factor