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Canadian journal of veterinary research = Revue canadienne de recherche veterinaire2016; 80(4); 294-301;

Evaluation of adipose-derived stromal vascular fraction from the lateral tailhead, inguinal region, and mesentery of horses.

Abstract: Use of mesenchymal stem cells (MSCs) found in the stromal vascular fraction (SVF) of equine adipose tissue has promising applications for regenerative therapies. The most commonly used source of equine adipose tissue is the subcutaneous tailhead. The objective of this study was to compare 3 adipose depot sites in horses and determine the viability and cellular yield, capillary density, gene expression for selected markers, and colony-forming unit fibroblasts (CFU-Fs) in adipose tissue taken from these sites. Adipose tissue was excised from the area lateral to the tailhead, the inguinal region, and the small colon mesentery of 6 horses. Lipoaspirate was also collected from the area lateral to the tailhead. Stromal vascular fraction (SVF) was prepared in duplicate from the 3 different adipose tissue depots. The total nucleated and dead cell counts was determined manually using a hemocytometer and percent viability was calculated. Mass and volume of adipose were determined in order to calculate density and factor-VIII immunohistochemical staining was used to determine vascular density in the excisional adipose tissue samples from each horse. Quantitative polymerase chain reaction (qPCR) was used to quantify gene expression for selected cellular markers from each site. There were significant differences in viability, yield of nucleated cells/gram of adipose tissue, vascular density, gene expression, and CFU-Fs among adipose depots. Adipose from the mesentery yielded the highest number of nucleated cells/gram of tissue and the highest vascular density and percentage of CFU-Fs. In the horse, both the anatomical site of collection and the method of tissue collection significantly impact the yield and composition of cells in the SVF. Further study is needed to assess whether one adipose source is superior for harvesting mesenchymal stem cells (MSCs) and whether the differences among sources are clinically relevant for treatment of musculoskeletal injuries in horses. L’utilisation de cellules souches mésenchymateuses (CSMs) retrouvées dans la fraction du stroma vasculaire (FSV) du tissu adipeux équin a des applications prometteuses pour les thérapies régénératrices. La source la plus fréquemment utilisée de tissu adipeux équin est le tissu sous-cutané de la base de la queue. L’objectif de la présente étude était de comparer trois sites de dépôts adipeux chez le cheval et de déterminer la viabilité et la récolte cellulaire, la densité capillaire, l’expression génique de marqueurs sélectionnés, et le nombre de fibroblastes formateur des colonies (FFC) dans le tissu adipeux prélevés de ces sites. Le tissu adipeux a été excisé de la région latérale à la base de la queue, de la région inguinale, et du mésentère du petit colon de six chevaux. Des aspirations de lipide ont également été prélevées de la région latérale de la base de la queue. La FSV a été préparée en duplicata à partir de chacun des trois dépôts différents de tissu adipeux. Les dénombrements totaux des cellules nucléées et mortes ont été déterminés manuellement à l’aide d’un hémocytomètre et le pourcentage de viabilité calculé. La masse et le volume de tissu adipeux ont été déterminés afin de calculer la densité et la coloration par immunohistochimie du facteur VIII a été utilisée afin de déterminer la densité vasculaire dans les échantillons de tissu adipeux excisé de chaque cheval. Une réaction d’amplification en chaine par la polymérase quantitative (ACPq) a été utilisée pour quantifier l’expression génique pour des marqueurs cellulaires sélectionnés de chaque site. Il y avait des différences significatives dans la viabilité, le rendement de cellules nucléées/gramme de tissu adipeux, la densité vasculaire, l’expression génique, et les FFCs entre les dépôts adipeux. Le tissu adipeux provenant du mésentère a généré le plus grand nombre de cellules nucléées/gramme de tissu et la plus haute densité vasculaire et pourcentage de FFCs. Chez le cheval, le site anatomique de prélèvement et la méthode de prélèvement du tissu ont un impact significatif sur le rendement et la composition cellulaire dans la FSV. Des études additionnelles sont requises pour évaluer si une source de tissu adipeux est supérieure pour récolter des cellules souches mésenchymateuses et si les différences entre les sources sont cliniquement pertinentes pour le traitement de blessures morpho-squelettiques chez les chevaux.(Traduit par Docteur Serge Messier).
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Publication Date: 2016-10-14 PubMed ID: 27733784PubMed Central: PMC5052881
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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 dives into the potential of using mesenchymal stem cells (MSCs) from horse adipose tissue for regenerative therapies. It examines three different areas of fat collection in the horse’s body and assesses the viability, cellular yield, capillary density, and gene expression, concluding that the method and location of the adipose extraction significantly influence the success of the procedure.

Objective and Methodology

  • The objective of the study was to analyze the stromal vascular fraction (SVF), a rich source of mesenchymal stem cells (MSCs), extracted from adipose tissues in different areas of six horses’ bodies.
  • The adipose tissues were collected from three distinct locations – the area lateral to the tailhead, the inguinal region, and the small colon mesentery.
  • Alongside excision, lipoaspiration was also performed on the area lateral to the tailhead.
  • The techniques used to evaluate the samples included quantitative polymerase chain reaction (qPCR) to assess gene expression, factor-VIII immunohistochemical staining to determine vascular density, and hemocytometer to evaluate total nucleated and dead cell counts.

Results and Findings

  • All adipose depot sites showed differing results in terms of viability, cellular yield, vascular density, gene expression, and percentage of colony-forming unit fibroblasts (CFU-Fs).
  • The adipose tissue from the mesentery yielded the highest number of nucleated cells per gram of tissue, the highest vascular density and the highest percentage of CFU-Fs, indicating it to be the most efficient site for MSCs extraction.

Impact and Recommendations

  • The results highlight the crucial role of the collection site and method in harvesting useful MSCs from the SVF of horse adipose tissues.
  • The findings could be significant for the development of new regenerative therapies focusing on stem cells derived from adipose tissues.
  • Further research should address if one adipose source is better than others in MSCs collection and study the potential clinical relevance in treating musculoskeletal injuries in horses.

Cite This Article

APA
Metcalf GL, McClure SR, Hostetter JM, Martinez RF, Wang C. (2016). Evaluation of adipose-derived stromal vascular fraction from the lateral tailhead, inguinal region, and mesentery of horses. Can J Vet Res, 80(4), 294-301.

Publication

Canadian journal of veterinary research = Revue canadienne de recherche veterinaire
ISSN: 1928-9022
NlmUniqueID: 8607793
Country: Canada
Language: English
Volume: 80
Issue: 4
Pages: 294-301

Researcher Affiliations

Metcalf, Garrett L
  • Department of Veterinary Clinical Sciences (Metcalf, McClure), Department of Pathology (Hostetter), Department of Diagnostics and Production Animal Medicine and Department of Statistics (Wang), Iowa State University, Ames, Iowa, USA; InGeneron Inc., 8205 El Rio, Houston, Texas, USA (Martinez).
McClure, Scott R
  • Department of Veterinary Clinical Sciences (Metcalf, McClure), Department of Pathology (Hostetter), Department of Diagnostics and Production Animal Medicine and Department of Statistics (Wang), Iowa State University, Ames, Iowa, USA; InGeneron Inc., 8205 El Rio, Houston, Texas, USA (Martinez).
Hostetter, Jesse M
  • Department of Veterinary Clinical Sciences (Metcalf, McClure), Department of Pathology (Hostetter), Department of Diagnostics and Production Animal Medicine and Department of Statistics (Wang), Iowa State University, Ames, Iowa, USA; InGeneron Inc., 8205 El Rio, Houston, Texas, USA (Martinez).
Martinez, Rudy F
  • Department of Veterinary Clinical Sciences (Metcalf, McClure), Department of Pathology (Hostetter), Department of Diagnostics and Production Animal Medicine and Department of Statistics (Wang), Iowa State University, Ames, Iowa, USA; InGeneron Inc., 8205 El Rio, Houston, Texas, USA (Martinez).
Wang, Chong
  • Department of Veterinary Clinical Sciences (Metcalf, McClure), Department of Pathology (Hostetter), Department of Diagnostics and Production Animal Medicine and Department of Statistics (Wang), Iowa State University, Ames, Iowa, USA; InGeneron Inc., 8205 El Rio, Houston, Texas, USA (Martinez).

MeSH Terms

  • Adipose Tissue / cytology
  • Animals
  • Cell Survival / physiology
  • Colony-Forming Units Assay
  • Fibroblasts / physiology
  • Gene Expression Regulation
  • Horses
  • Immunohistochemistry
  • Mesenchymal Stem Cells / physiology
  • Tissue and Organ Harvesting / veterinary

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Citations

This article has been cited 9 times.
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