Abstract
BackgroundInduced pluripotent stem (iPS) cells are generated from mouse and human somatic cells by the forced expression of defined transcription factors. Although most somatic cells are capable of acquiring pluripotency with minimal gene transduction, the poor efficiency of cell reprogramming and the uneven quality of iPS cells are still important problems. In particular, the choice of cell type most suitable for inducing high-quality iPS cells remains unclear.Methodology/Principal FindingsHere, we generated iPS cells from PDGFRα+ Sca-1+ (PαS) adult mouse mesenchymal stem cells (MSCs) and PDGFRα− Sca-1− osteo-progenitors (OP cells), and compared the induction efficiency and quality of individual iPS clones. MSCs had a higher reprogramming efficiency compared with OP cells and Tail Tip Fibroblasts (TTFs). The iPS cells induced from MSCs by Oct3/4, Sox2, and Klf4 appeared to be the closest equivalent to ES cells by DNA microarray gene profile and germline-transmission efficiency.Conclusions/SignificanceOur findings suggest that a purified source of undifferentiated cells from adult tissue can produce high-quality iPS cells. In this context, prospectively enriched MSCs are a promising candidate for the efficient generation of high-quality iPS cells.
Highlights
Pioneering work by Takahashi et al showed that the ectopic expression of a defined set of transcription factors, Oct4, Klf4, Sox2, and c-Myc, reprograms mouse embryonic fibroblasts (MEFs) and adult tail-tip fibroblasts (TTFs) into embryonic stem (ES)-like cells called induced pluripotent stem cells [1]
Each isolated cell type (PaS, OP, Tail Tip Fibroblast (TTF), and Mouse Embryonic Fibroblast (MEF)) was retrovirally transduced with 4 or 3 factors [15] along with CAG-DsRed [16] as a control for the induction efficiency, which was similar for both cell populations (Figure S1)
From what we found in the comparison of GFP+/DsRed2 colonies was that the PDGFRa+ Sca-1+ (PaS)
Summary
Pioneering work by Takahashi et al showed that the ectopic expression of a defined set of transcription factors, Oct, Klf, Sox, and c-Myc, reprograms mouse embryonic fibroblasts (MEFs) and adult tail-tip fibroblasts (TTFs) into embryonic stem (ES)-like cells called induced pluripotent stem (iPS) cells [1]. IPS cells have been generated from a variety of somatic cells, including embryonic and adult dermal fibroblasts [1,2,3], epithelial cells of the liver and stomach [4], pancreatic b cells [5], mature B lymphocytes [6], and adult neural stem cells (NSCs) [7,8]. These studies demonstrated that most somatic cells can be reprogrammed with 4 or 3 factors (excluding c-Myc). The choice of cell type most suitable for inducing high-quality iPS cells remains unclear
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