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  • Enhanced generation of induced pluripotent stem cells from a subpopulation of human fibroblasts.

Enhanced generation of induced pluripotent stem cells from a subpopulation of human fibroblasts.

PloS one (2009-09-24)
James A Byrne, Ha Nam Nguyen, Renee A Reijo Pera
ABSTRACT

The derivation of induced pluripotent stem cells (iPSCs) provides new possibilities for basic research and novel cell-based therapies. Limitations, however, include our current lack of understanding regarding the underlying mechanisms and the inefficiency of reprogramming. Here, we report identification and isolation of a subpopulation of human dermal fibroblasts that express the pluripotency marker stage specific embryonic antigen 3 (SSEA3). Fibroblasts that expressed SSEA3 demonstrated an enhanced iPSC generation efficiency, while no iPSC derivation was obtained from the fibroblasts that did not express SSEA3. Transcriptional analysis revealed NANOG expression was significantly increased in the SSEA3 expressing fibroblasts, suggesting a possible mechanistic explanation for the differential reprogramming. To our knowledge, this study is the first to identify a pluripotency marker in a heterogeneous population of human dermal fibroblasts, to isolate a subpopulation of cells that have a significantly increased propensity to reprogram to pluripotency and to identify a possible mechanism to explain this differential reprogramming. This discovery provides a method to significantly increase the efficiency of reprogramming, enhancing the feasibility of the potential applications based on this technology, and a tool for basic research studies to understand the underlying reprogramming mechanisms.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-TRA-1-60 Antibody, clone TRA-1-60, clone TRA-1-60, Chemicon®, from mouse
Sigma-Aldrich
Anti-Stage-Specific Embryonic Antigen-4 Antibody, clone MC-813-70, clone MC-813-70, Chemicon®, from mouse