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Stem cells international2012; 2012; 429160; doi: 10.1155/2012/429160

Induction of pluripotency in adult equine fibroblasts without c-MYC.

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.
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Publication Date: 2012-03-19 PubMed ID: 22550508PubMed Central: PMC3328202DOI: 10.1155/2012/429160Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article centers on the successful induction of pluripotency in adult equine fibroblasts, a significant step in equine embryonic stem cell research. The study demonstrates that pluripotency can be achieved using three transcription factors, OCT4, SOX2, and KLF4 without the c-MYC factor.

Objective and Methodology

  • The objective of this research was to induce pluripotency in adult equine fibroblasts, an important companion species, by sidestepping the use of the c-MYC transcription factor, believed until now to be essential.
  • To achieve this, the fibroblasts were transducted via retroviral means with three transcription factors namely OCT4, SOX2, and KLF4.

Results and Findings

  • The successfully induced pluripotent stem (iPS) cell lines, named EiPS, were maintained for over 27 passages that is over a period of 11 months.
  • The EiPS cells stained positive for alkaline phosphatase, a signal of pluripotency.
  • Further characterization of the EiPS cells led to the observation that they expressed OCT4, NANOG, SSEA1, and SSEA4, essential makers of pluripotency.
  • A gene analysis revealed the expression of OCT4, SOX2, NANOG, and STAT3, strengthening the proof of pluripotency achieved through the used methods.
  • The EiPS cells preserved their normal chromosome count of 64 even after long-term culture cryopreservation.

Conclusive Remarks

  • A key conclusion was the demonstrative proof that EiPS cells possess the capacity for differentiation into three embryonic germ layers, validated both in vitro by producing embryoid bodies and in vivo by creating teratomas.
  • This significant achievement in equine embryonic stem cell research, the derivation of iPS cells from adult equine fibroblasts, can be traced back to the use of the three reprogramming factors along with long-term maintenance suggesting sustained pluripotency.

Cite This Article

APA
Khodadadi K, Sumer H, Pashaiasl M, Lim S, Williamson M, Verma PJ. (2012). Induction of pluripotency in adult equine fibroblasts without c-MYC. Stem Cells Int, 2012, 429160. https://doi.org/10.1155/2012/429160

Publication

Stem cells international
ISSN: 1687-9678
NlmUniqueID: 101535822
Country: United States
Language: English
Volume: 2012
Pages: 429160
PII: 429160

Researcher Affiliations

Khodadadi, Khodadad
  • Centre for Reproduction and Development, Monash Institute of Medical Research, Monash University, Clayton, VIC 3800, Australia.
Sumer, Huseyin
    Pashaiasl, Maryam
      Lim, Susan
        Williamson, Mark
          Verma, Paul J

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