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Stem Cells International
Volume 2012, Article ID 429160, 9 pages
http://dx.doi.org/10.1155/2012/429160
Research Article

Induction of Pluripotency in Adult Equine Fibroblasts without c-MYC

1Centre for Reproduction and Development, Monash Institute of Medical Research, Monash University, Clayton, VIC 3800, Australia
2Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz 51666-14766, Iran
3Stem Cell Technologies i (SCTi), Gleneagles Medical Centre, Singapore 258499
4Gribbles Veterinary, Clayton, VIC 3168, Australia
5South Australian Research Institute (SARDI), Turretfield Research Centre, Rosedale, SA 5350, Australia

Received 9 November 2011; Revised 28 December 2011; Accepted 3 January 2012

Academic Editor: Rajarshi Pal

Copyright © 2012 Khodadad Khodadadi et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Despite tremendous efforts on isolation of pluripotent equine embryonic stem (ES) cells, to date there are few reports about successful isolation of ESCs and no report of in vivo differentiation of this important companion species. We report the induction of pluripotency in adult equine fibroblasts via retroviral transduction with three transcription factors using OCT4, SOX2, and KLF4 in the absence of c-MYC. The cell lines were maintained beyond 27 passages (more than 11 months) and characterized. The equine iPS (EiPS) cells stained positive for alkaline phosphatase by histochemical staining and expressed OCT4, NANOG, SSEA1, and SSEA4. Gene expression analysis of the cells showed the expression of OCT4, SOX2 NANOG, and STAT3. The cell lines retained a euploid chromosome count of 64 after long-term culture cryopreservation. The EiPS demonstrated differentiation capacity for the three embryonic germ layers both in vitro by embryoid bodies (EBs) formation and in vivo by teratoma formation. In conclusion, we report the derivation of iPS cells from equine adult fibroblasts and long-term maintenance using either of the three reprogramming factors.