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Home / Equine Research Bank / Stem Cell Res Ther / Isolation and characterization of equine native MSC populati...
Stem cell research & therapy2017; 8(1); 80; doi: 10.1186/s13287-017-0525-2

Isolation and characterization of equine native MSC populations.

Abstract: In contrast to humans in which mesenchymal stem/stromal cell (MSC) therapies are still largely in the clinical trial phase, MSCs have been used therapeutically in horses for over 15 years, thus constituting a valuable preclinical model for humans. In human tissues, MSCs have been shown to originate from perivascular cells, namely pericytes and adventitial cells, which are identified by the presence of the cell surface markers CD146 and CD34, respectively. In contrast, the origin of MSCs in equine tissues has not been established, preventing the isolation and culture of defined cell populations in that species. Moreover, a comparison between perivascular CD146 and CD34 cell populations has not been performed in any species. Immunohistochemistry was used to identify adventitial cells (CD34) and pericytes (CD146) and to determine their localization in relation to MSCs in equine tissues. Isolation of CD34 (CD34/CD146/CD144/CD45) and CD146 (CD146/CD34/CD144/CD45) cell fractions from equine adipose tissue was achieved by fluorescence-activated cell sorting. The isolated cell fractions were cultured and analyzed for the expression of MSC markers, using qPCR and flow cytometry, and for the ability to undergo trilineage differentiation. Angiogenic properties were analyzed in vivo using a chorioallantoic membrane (CAM) assay. Both CD34 and CD146 cells displayed typical MSC features, namely growth in uncoated tissue culture dishes, clonal growth when seeded at low density, expression of typical MSC markers, and multipotency shown by the capacity for trilineage differentiation. Of note, CD146 cells were distinctly angiogenic compared with CD34 and non-sorted cells (conventional MSCs), demonstrated by the induction of blood vessels in a CAM assay, expression of elevated levels of VEGFA and ANGPT1, and association with vascular networks in cocultures with endothelial cells, indicating that CD146 cells maintain a pericyte phenotype in culture. This study reports for the first time the successful isolation and culture of CD146 and CD34 cell populations from equine tissues. Characterization of these cells evidenced their distinct properties and MSC-like phenotype, and identified CD146 cells as distinctly angiogenic, which may provide a novel source for enhanced regenerative therapies.
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Publication Date: 2017-04-18 PubMed ID: 28420427PubMed Central: PMC5395828DOI: 10.1186/s13287-017-0525-2Google 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 article discusses a research that studies the origin of Mesenchymal Stem/Stromal Cells (MSCs) in equines or horses. The researchers have successfully identified and characterised equine MSCs that originate from perivascular cells, a feature previously unidentified in equines. Notably, they found that CD146 cells, a type of perivascular cell, are distinctly angiogenic, suggesting possible applications in improved regenerative therapies.

Objective and Methodology

  • This research aimed to understand the origin of mesenchymal stem/stromal cells (MSCs) in horses, which was unknown till now. This is important to enable the isolation and culture of defined cell populations in horses.
  • The research methodology involved the use of immunohistochemistry to identify perivascular cells, specifically pericytes (CD146) and adventitial cells (CD34) in horse tissues. These cells were compared for the first time in any species.
  • Fluorescence-activated cell sorting was used to isolate CD34 and CD146 cell fractions from equine adipose tissue.
  • The isolated cell fractions were cultured and analysed for the expression of MSC marker cells, their capacity for trilineage differentiation, and angiogenic properties.

Results and Findings

  • The research found that both CD34 and CD146 cells displayed typical characteristics of MSCs, including growth in uncoated tissue culture dishes, clonal growth when seeded at low density, and expression of typical MSC markers. They also showed multipotency – the capacity for trilineage differentiation.
  • Significantly, CD146 cells were found to be distinctly angiogenic compared to CD34 and non-sorted cells (conventional MSCs). Angiogenic cells stimulate the growth of new blood vessels.
  • The angiogenic nature of CD146 cells was demonstrated by the induction of blood vessels in a chorioallantoic membrane (CAM) assay, expression of elevated levels of VEGFA and ANGPT1, and association with vascular networks in cocultures with endothelial cells. This indicates that CD146 cells maintain a pericyte phenotype in culture.

Significance and Implications

  • This research constitutes the first successful isolation and culture of CD146 and CD34 cell populations from equine tissues.
  • The distinct properties and MSC-like phenotype of these cells were characterized and CD146 cells identified as distinctly angiogenic.
  • The angiogenic CD146 cells could provide a novel source for enhanced regenerative therapies, thus having significant implications for veterinary medicine and potential parallels in human medicine.

Cite This Article

APA
Esteves CL, Sheldrake TA, Mesquita SP, Pesántez JJ, Menghini T, Dawson L, Péault B, Donadeu FX. (2017). Isolation and characterization of equine native MSC populations. Stem Cell Res Ther, 8(1), 80. https://doi.org/10.1186/s13287-017-0525-2

Publication

Stem cell research & therapy
ISSN: 1757-6512
NlmUniqueID: 101527581
Country: England
Language: English
Volume: 8
Issue: 1
Pages: 80
PII: 80

Researcher Affiliations

Esteves, Cristina L
  • The Roslin Institute, University of Edinburgh, Edinburgh, UK. cristina.esteves@roslin.ed.ac.uk.
  • The Roslin Institute, University of Edinburgh, Easter Bush, Midlothian, EH25 9RG, UK. cristina.esteves@roslin.ed.ac.uk.
Sheldrake, Tara A
  • The Roslin Institute, University of Edinburgh, Edinburgh, UK.
Mesquita, Simone P
  • The Roslin Institute, University of Edinburgh, Edinburgh, UK.
Pesántez, Juan J
  • The Roslin Institute, University of Edinburgh, Edinburgh, UK.
Menghini, Timothy
  • The Roslin Institute, University of Edinburgh, Edinburgh, UK.
Dawson, Lucy
  • The Roslin Institute, University of Edinburgh, Edinburgh, UK.
Péault, Bruno
  • Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, UK.
  • Orthopaedic Hospital Research Centre, University of California, Los Angeles, CA, USA.
Donadeu, F Xavier
  • The Roslin Institute, University of Edinburgh, Edinburgh, UK.

MeSH Terms

  • Adipose Tissue / cytology
  • Angiopoietin-1 / genetics
  • Angiopoietin-1 / metabolism
  • Animals
  • Antigens, CD34 / genetics
  • Antigens, CD34 / metabolism
  • CD146 Antigen / genetics
  • CD146 Antigen / metabolism
  • Cells, Cultured
  • Horses
  • Mesenchymal Stem Cells / cytology
  • Mesenchymal Stem Cells / metabolism
  • Pericytes / cytology
  • Pericytes / metabolism
  • Primary Cell Culture / veterinary
  • Vascular Endothelial Growth Factor A / genetics
  • Vascular Endothelial Growth Factor A / metabolism

Grant Funding

  • G1000816 / Medical Research Council

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