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Home / Equine Research Bank / In Vitro Cell Dev Biol Anim / Immunophenotypic characterization and tenogenic differentiat...
In vitro cellular & developmental biology. Animal2014; 50(6); 538-548; doi: 10.1007/s11626-013-9729-7

Immunophenotypic characterization and tenogenic differentiation of mesenchymal stromal cells isolated from equine umbilical cord blood.

Abstract: Mesenchymal stem cells (MSCs) isolated from umbilical cord blood (UCB) in equines have not been well characterized with respect to the expression of pluripotency and mesenchymal markers and for tenogenic differentiation potential in vitro. The plastic adherent fibroblast-like cells isolated from 13 out of 20 UCB samples could proliferate till passage 20. The cells expressed pluripotency markers (OCT4, NANOG, and SOX2) and MSC surface markers (CD90, CD73, and CD105) by RT-PCR, but did not express CD34, CD45, and CD14. On immunocytochemistry, the isolated cells showed expression of CD90 and CD73 proteins, but tested negative for CD34 and CD45. In flow cytometry, CD29, CD44, CD73, and CD90 were expressed by 96.36 ± 1.28%, 93.40 ± 0.70%, 73.23 ± 1.29% and 46.75 ± 3.95% cells, respectively. The UCB-MSCs could be differentiated to tenocytes by culturing in growth medium supplemented with 50 ng/ml of BMP-12 by day 10. The differentiated cells showed the expression of mohawk homeobox (Mkx), collagen type I alpha 1 (Col1α1), scleraxis (Scx), tenomodulin (Tnmd) and decorin (Dcn) by RT-PCR. In addition, flow cytometry detected tenomodulin and decorin protein in 95.65 ± 2.15% and 96.30 ± 1.00% of differentiated cells in comparison to 11.30 ± 0.10% and 19.45 ± 0.55% cells, respect vely in undifferentiated control cells. The findings support the observation that these cells may be suitable for therapeutic applications, including ruptured tendons in racehorses.
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Publication Date: 2014-01-11 PubMed ID: 24414976DOI: 10.1007/s11626-013-9729-7Google 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.

The research explores the properties of mesenchymal stem cells (MSCs) derived from equine umbilical cord blood, particularly their potential for differentiation into tendon cells (tenocytes), which could have therapeutic applications in treating tendon injuries in horses.

Overview of the Research

  • The researchers obtained MSCs from the umbilical cord blood of horses. These cells were isolated and placed in in-vitro conditions where they were allowed to proliferate and grow until passage 20.
  • A total of 20 umbilical cord blood samples were used for this study, with cells successfully isolated from 13 of them. This suggests a successful isolation rate of about 65%.
  • The isolated cells showed fibroblast-like characteristics (cells that produce collagen and other fibres), which is typical of MSCs.

Characterization of Cells

  • The cells were tested for the expression of certain markers typical of MSCs and cells in a state of pluripotency – the potential to develop into any cell type.
  • It was found that they expressed pluripotency markers OCT4, NANOG, and SOX2, and MSC surface markers CD90, CD73, and CD105. However, they did not express markers CD34, CD45, and CD14.
  • These findings were validated through RT-PCR techniques, immunocytochemistry, and flow cytometry – all processes that reliably help detect the presence of specific proteins or markers in cells.

Cell Differentiation

  • The researchers then attempted to differentiate these cells into tenocytes (tendon cells), by culturing them in growth medium supplemented with a protein called BMP-12.
  • By day 10 of culturing, these stem cells showed the expression of certain markers indicative of tenocytes, including mohawk homeobox (Mkx), collagen type I alpha 1 (Col1α1), scleraxis (Scx), tenomodulin (Tnmd), and decorin (Dcn), which were confirmed through RT-PCR and flow cytometry.
  • High percentages of tenomodulin and decorin proteins were detected in the differentiated cells, suggesting successful tenogenic differentiation.

Relevance to Therapy

  • The research supports the potential use of these cells in therapeutic applications. Specifically, they could be used to aid in the repair of ruptured tendons in racehorses, a common ailment in the racing industry.
  • The ability to encourage MSCs derived from a horse’s umbilical cord blood to differentiate into tenocytes could potentially offer a new therapy for equine tendon injuries. More research is needed to confirm efficacy and safety.

Cite This Article

APA
Mohanty N, Gulati BR, Kumar R, Gera S, Kumar P, Somasundaram RK, Kumar S. (2014). Immunophenotypic characterization and tenogenic differentiation of mesenchymal stromal cells isolated from equine umbilical cord blood. In Vitro Cell Dev Biol Anim, 50(6), 538-548. https://doi.org/10.1007/s11626-013-9729-7

Publication

In vitro cellular & developmental biology. Animal
ISSN: 1543-706X
NlmUniqueID: 9418515
Country: Germany
Language: English
Volume: 50
Issue: 6
Pages: 538-548

Researcher Affiliations

Mohanty, Niharika
  • Department of Veterinary Physiology and Biochemistry, College of Veterinary Sciences, LLR University of Veterinary & Animal Sciences, Hisar, 25004, Haryana, India.
Gulati, Baldev R
    Kumar, Rajesh
      Gera, Sandeep
        Kumar, Pawan
          Somasundaram, Rajesh K
            Kumar, Sandeep

              MeSH Terms

              • 5'-Nucleotidase / biosynthesis
              • Animals
              • Cell Differentiation / physiology
              • Cells, Cultured
              • Decorin / biosynthesis
              • Fetal Blood / cytology
              • Homeodomain Proteins / biosynthesis
              • Horses
              • Membrane Proteins / biosynthesis
              • Mesenchymal Stem Cells / cytology
              • Mesenchymal Stem Cells / metabolism
              • Octamer Transcription Factor-3 / biosynthesis
              • SOXB1 Transcription Factors / biosynthesis
              • Thy-1 Antigens / biosynthesis

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