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Home / Equine Research Bank / Can J Vet Res / Application of a novel sorting system for equine mesenchymal...
Canadian journal of veterinary research = Revue canadienne de recherche veterinaire2014; 78(4); 290-296;

Application of a novel sorting system for equine mesenchymal stem cells (MSCs).

Abstract: The objective of this study was to validate non-equilibrium gravitational field-flow fractionation (GrFFF), an immunotag-less method of sorting mesenchymal stem cells (MSCs) into subpopulations, for use with MSCs derived from equine muscle tissue, periosteal tissue, bone marrow, and adipose tissue. Cells were collected from 6 young, adult horses, postmortem. Cells were isolated from left semitendinosus muscle tissue, periosteal tissue from the distomedial aspect of the right tibia, bone marrow aspirates from the fourth and fifth sternebrae, and left supragluteal subcutaneous adipose tissue. Aliquots of 800 × 10(3) MSCs from each tissue source were separated and injected into a ribbon-like capillary device by continuous flow (GrFFF proprietary system). Cells were sorted into 6 fractions and absorbencies [optical density (OD)] were read. Six fractions from each of the 6 aliquots were then combined to provide pooled fractions that had adequate cell numbers to seed at equal concentrations into assays. Equine muscle tissue-derived, periosteal tissue-derived, bone marrow-derived, and adipose tissue-derived mesenchymal stem cells were consistently sorted into 6 fractions that remained viable for use in further assays. Fraction 1 had more cuboidal morphology in culture when compared to the other fractions. Statistical analysis of the fraction absorbencies (OD) revealed a P-value of < 0.05 when fractions 2 and 3 were compared to fractions 1, 4, 5, and 6. It was concluded that non-equilibrium GrFFF is a valid method for sorting equine muscle tissue-derived, periosteal tissue-derived, bone marrow-derived, and adipose tissue-derived mesenchymal stem cells into subpopulations that remain viable, thus securing its potential for use in equine stem cell applications and veterinary medicine. L’objectif de la présente étude était de valider une méthode non-équilibrée de fractionnement par flot sous champs gravitationnel (GrFFF), une méthode sans marquage immunologique de séparation des cellules souches mésenchymateuses (MSCs) en sous-populations, pour utilisations avec des MSCs provenant de tissu musculaire, de tissu de périoste, de moelle osseuse, et de tissu adipeux de chevaux. Les cellules furent prélevées post-mortem à partir de six jeunes chevaux adultes. Les cellules furent isolées du muscle semi-tendineux gauche, du périoste de l’aspect disto-médial du tibia droit, d’aspirations de moelle osseuse de la quatrième et cinquième sternèbres, et du tissu adipeux sous-cutané de la région supra-glutéale gauche. Des aliquots de 800 × 10 MSCs de chaque tissu ont été séparés et injectés dans un appareil capillaire apparenté à un ruban par flot continu (système breveté GrFFF). Les cellules furent séparées en six fractions et les absorbances [densité optique (OD)] notées. Six fractions de chacun des six aliquots furent par la suite combinées afin de fournir des fractions poolées qui avaient des nombres adéquats de cellules pour ensemencer des concentrations égales dans les essais. Les MSCs provenant du tissu musculaire, du périoste, de la moelle osseuse, et du tissu adipeux étaient de manière constante séparées en six fractions qui sont demeurées viables pour utilisation dans des essais ultérieurs. La fraction 1 avait plus de cellules de morphologie cuboïde comparativement aux autres fractions. Les analyses statistiques des OD des fractions ont révélé une valeur de < 0,05 lorsque les fractions 2 et 3 étaient comparées aux fractions 1, 4, 5, et 6. Il fut conclu que la méthode GrFFF non-équilibrée est une méthode valide pour séparer les MSCs équines dérivées des cellules musculaires, du périoste, de la moelle osseuse, et du tissu adipeux en sous-populations qui demeurent viables, assurant ainsi son potentiel pour utilisation en médecine vétérinaire et les applications avec les cellules souches équines.(Traduit par Docteur Serge Messier).
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Publication Date: 2014-10-31 PubMed ID: 25355998PubMed Central: PMC4170768
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't
  • Validation Study

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 study examines the effectiveness of a unique sorting system known as non-equilibrium GrFFF in identifying and categorizing mesenchymal stem cells (MSCs) from various horse tissues, such as muscle, bone marrow, periosteal, and adipose for use in veterinary medicine and horse stem cell applications.

Research Approach

  • The study identifies the different sources of MSC’s which include muscle, bone marrow, periosteal and adipose tissues from six young horses.
  • These horses were studied postmortem and cells were specifically isolated from different parts and tissues such as the left semitendinosus (a muscle in the posterior thigh), periosteal tissue from the distomedial aspect of the right tibia, bone marrow aspirates from the fourth and fifth sternebrae, and left supragluteal subcutaneous adipose tissue.
  • Each cell source was divided into small volumes of 800 × 10(3) MSCs and were then processed by the GrFFF system.
  • The GrFFF system executed its function by introducing the cells into a narrow, ribbon-like capillary device. The functionality of this is to isolate and sort cells into sub-groups or fractions.

Results and Interpretations

  • As a result, the MSCs from each tissue source could be consistently categorized into 6 fractions. These fractions were then combined to form pooled fractions that held viable cell numbers.
  • The study reported that the fraction one, of all the sub-divisions, had a more ‘cuboidal’ morphology when compared to the other fractions.
  • Performing a statistical analysis, it was determined that fractions 2 and 3 showed significant differences in optical density as compared to fractions 1, 4, 5, and 6 which indicates a different cell distribution within these fractions.

Conclusion

  • The study concluded that the non-equilibrium GrFFF is a valid method for categorizing horse tissue-derived MSCs, keeping them viable for future use.
  • This discovery strengthens the potential of this method for use in horse stem cell applications or veterinary medicine in general.

Cite This Article

APA
Radtke CL, Nino-Fong R, Esparza Gonzalez BP, McD￮ LA. (2014). Application of a novel sorting system for equine mesenchymal stem cells (MSCs). Can J Vet Res, 78(4), 290-296.

Publication

Canadian journal of veterinary research = Revue canadienne de recherche veterinaire
ISSN: 1928-9022
NlmUniqueID: 8607793
Country: Canada
Language: English
Volume: 78
Issue: 4
Pages: 290-296

Researcher Affiliations

Radtke, Catherine L
  • Department of Health Management, Atlantic Veterinary College, University of Prince Edward Island, 550 University Avenue, Charlottetown, Prince Edward Island C1A 4P3.
Nino-Fong, Rodolfo
  • Department of Health Management, Atlantic Veterinary College, University of Prince Edward Island, 550 University Avenue, Charlottetown, Prince Edward Island C1A 4P3.
Esparza Gonzalez, Blanca P
  • Department of Health Management, Atlantic Veterinary College, University of Prince Edward Island, 550 University Avenue, Charlottetown, Prince Edward Island C1A 4P3.
McD￮, Laurie A
  • Department of Health Management, Atlantic Veterinary College, University of Prince Edward Island, 550 University Avenue, Charlottetown, Prince Edward Island C1A 4P3.

MeSH Terms

  • Adipose Tissue / cytology
  • Animals
  • Cell Differentiation / physiology
  • Fractionation, Field Flow / instrumentation
  • Fractionation, Field Flow / methods
  • Horses
  • Mesenchymal Stem Cells / cytology
  • Muscle, Skeletal / cytology
  • Periosteum / cytology
  • Tibia / cytology

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Citations

This article has been cited 3 times.
  1. Radtke CL, Nino-Fong R, Rodriguez-Lecompte JC, Esparza Gonzalez BP, Stryhn H, McD￮ LA. Osteogenic potential of sorted equine mesenchymal stem cell subpopulations. Can J Vet Res 2015 Apr;79(2):101-8.
    pubmed: 25852225
  2. Ferreira-Baptista C, Ferreira R, Fernandes MH, Gomes PS, Colaço B. Influence of the Anatomical Site on Adipose Tissue-Derived Stromal Cells' Biological Profile and Osteogenic Potential in Companion Animals. Vet Sci 2023 Nov 24;10(12).
    doi: 10.3390/vetsci10120673pubmed: 38133224google scholar: lookup
  3. Feng X, Qi F, Wang H, Li W, Gan Y, Qi C, Lin Z, Chen L, Wang P, Hu Z, Miao Y. Sorting Technology for Mesenchymal Stem Cells from a Single Tissue Source. Stem Cell Rev Rep 2024 Feb;20(2):524-537.
    doi: 10.1007/s12015-023-10635-wpubmed: 38112926google scholar: lookup
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