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Stem cells and development2012; 22(4); 611-621; doi: 10.1089/scd.2012.0052

Derivation and characterization of induced pluripotent stem cells from equine fibroblasts.

Abstract: Pluripotent stem cells offer unprecedented potential not only for human medicine but also for veterinary medicine, particularly in relation to the horse. Induced pluripotent stem cells (iPSCs) are particularly promising, as they are functionally similar to embryonic stem cells and can be generated in vitro in a patient-specific manner. In this study, we report the generation of equine iPSCs from skin fibroblasts obtained from a foal and reprogrammed using viral vectors coding for murine Oct4, Sox2, c-Myc, and Klf4 sequences. The reprogrammed cell lines were morphologically similar to iPSCs reported from other species and could be stably maintained over more than 30 passages. Immunostaining and polymerase chain reaction analyses revealed that these cell lines expressed an array of endogenous markers associated with pluripotency, including OCT4, SOX2, NANOG, REX1, LIN28, SSEA1, SSEA4, and TRA1-60. Furthermore, under the appropriate conditions, the equine iPSCs readily formed embryoid bodies and differentiated in vitro into cells expressing markers of ectoderm, mesoderm, and endoderm, and when injected into immunodeficient mice, gave raise to tumors containing differentiated derivatives of the 3 germ layers. Finally, we also reprogrammed fibroblasts from a 2-year-old horse. The reprogrammed cells were similar to iPSCs derived from neonatal fibroblasts in terms of morphology, expression of pluripotency markers, and differentiation ability. The generation of these novel cell lines constitutes an important step toward the understanding of pluripotency in the horse, and paves the way for iPSC technology to potentially become a powerful research and clinical tool in veterinary biomedicine.
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Publication Date: 2012-09-28 PubMed ID: 22897112PubMed Central: PMC3564467DOI: 10.1089/scd.2012.0052Google Scholar: Lookup
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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.

This research study focuses on the development of pluripotent stem cells from horse fibroblasts, which could be a significant step in veterinary biomedicine. The creation of these equine induced pluripotent stem cells (iPSCs) was achieved through reprogramming methods, using specific sequences to help form reprogrammed cell lines. The study showed that these cell lines were able to differentiate into various cell types, thus demonstrating their pluripotency.

Generation of equine induced Pluripotent Stem Cells

  • The researchers generated equine iPSCs from skin fibroblasts taken from a young horse, using viral vectors coded with specific sequences (Oct4, Sox2, c-Myc, and Klf4) found in murine (mice).
  • The cells were reprogrammed to form cell lines, which maintained their stability for more than 30 passages. This high level of stability is a critical factor for clinical or lab use.

Verification of Pluripotency markers

  • The equine iPSCs demonstrated the expression of various pluripotency markers including OCT4, SOX2, NANOG, REX1, LIN28, SSEA1, SSEA4, and TRA1-60, which were confirmed by immunostaining and polymerase chain reaction (PCR) analyses.
  • The expression of these endogenous markers is a good indicator of the pluripotency of these reprogrammed cell lines.

Differentiation of equine iPSCs

  • The research further assessed the differentiation ability of these equine iPSCs by forming embryoid bodies and differentiating into cells expressing markers of ectoderm, mesoderm, and endoderm under suitable conditions.
  • The cells also formed tumors, comprising differentiated derivatives of the three germ layers when injected into immunodeficient mice, further corroborating the pluripotency of these cells.

Reprogramming of fibroblasts from an older horse

  • In addition to fibroblasts from a young horse, the researchers also reprogrammed fibroblasts from a two-year-old horse.
  • The cells obtained were as pluripotent as those from the young horse in terms of morphology, marker expression, and differentiation ability, which again solidifies the robustness of the reprogramming process implemented in this study.

Implications for Veterinary Biomedicine

  • This study signifies a crucial step towards understanding pluripotency in horses and could represent a powerful research tool in the field of veterinary biomedicine.
  • The creation of these equine iPSCs paves the way for this technology to become potentially valuable for treating various equine diseases or for developing new therapies in the field of veterinary medicine.

Cite This Article

APA
Breton A, Sharma R, Diaz AC, Parham AG, Graham A, Neil C, Whitelaw CB, Milne E, Donadeu FX. (2012). Derivation and characterization of induced pluripotent stem cells from equine fibroblasts. Stem Cells Dev, 22(4), 611-621. https://doi.org/10.1089/scd.2012.0052

Publication

Stem cells and development
ISSN: 1557-8534
NlmUniqueID: 101197107
Country: United States
Language: English
Volume: 22
Issue: 4
Pages: 611-621

Researcher Affiliations

Breton, Amandine
  • The Roslin Institute, University of Edinburgh, Midlothian, United Kingdom.
Sharma, Ruchi
    Diaz, Andrea Catalina
      Parham, Alea Gillian
        Graham, Audrey
          Neil, Claire
            Whitelaw, Christopher Bruce
              Milne, Elspeth
                Donadeu, Francesc Xavier

                  MeSH Terms

                  • Animals
                  • Antigens, Differentiation / biosynthesis
                  • Antigens, Differentiation / genetics
                  • Cell Dedifferentiation
                  • Cells, Cultured
                  • Embryoid Bodies / cytology
                  • Embryoid Bodies / metabolism
                  • Fibroblasts / cytology
                  • Fibroblasts / metabolism
                  • Horses
                  • Humans
                  • Induced Pluripotent Stem Cells / cytology
                  • Induced Pluripotent Stem Cells / metabolism
                  • Kruppel-Like Factor 4
                  • Mice
                  • Transcription Factors / biosynthesis
                  • Transcription Factors / genetics

                  Grant Funding

                  • BBS/E/D/05251442 / Biotechnology and Biological Sciences Research Council
                  • BBS/E/D/05251443 / Biotechnology and Biological Sciences Research Council
                  • BBS/E/D/05251444 / Biotechnology and Biological Sciences Research Council
                  • BBS/E/D/05251445 / Biotechnology and Biological Sciences Research Council

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                  This article has been cited 35 times.
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