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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

References

This article includes 26 references
  1. Triffitt JT. The stem cell of the osteoblast. In: Bilezikian JP, Raisz LG, Rodan GA, editors. Principles of Bone Biology. Vol. 1. San Diego, California: Academic Press; 1996. pp. 39–50.
  2. Radtke CL, Nino-Fong R, Esparza Gonzalez BP, Stryhn H, McD￮ LA. Characterization and osteogenic potential of equine muscle tissue- and periosteal tissue-derived mesenchymal stem cells in comparison with bone marrow- and adipose tissue-derived mesenchymal stem cells.. Am J Vet Res 2013 May;74(5):790-800.
    pubmed: 23627394doi: 10.2460/ajvr.74.5.790google scholar: lookup
  3. Roda B, Lanzoni G, Alviano F, Zattoni A, Costa R, Di Carlo A, Marchionni C, Franchina M, Ricci F, Tazzari PL, Pagliaro P, Scalinci SZ, Bonsi L, Reschiglian P, Bagnara GP. A novel stem cell tag-less sorting method.. Stem Cell Rev Rep 2009 Dec;5(4):420-7.
    pubmed: 20058204doi: 10.1007/s12015-009-9088-7google scholar: lookup
  4. Roda B, Zattoni A, Reschiglian P, Moon MH, Mirasoli M, Michelini E, Roda A. Field-flow fractionation in bioanalysis: A review of recent trends.. Anal Chim Acta 2009 Mar 9;635(2):132-43.
    pubmed: 19216870doi: 10.1016/j.aca.2009.01.015google scholar: lookup
  5. De Schauwer C, Meyer E, Van de Walle GR, Van Soom A. Markers of stemness in equine mesenchymal stem cells: a plea for uniformity.. Theriogenology 2011 May;75(8):1431-43.
    pubmed: 21196039doi: 10.1016/j.theriogenology.2010.11.008google scholar: lookup
  6. Taylor SE, Smith RK, Clegg PD. Mesenchymal stem cell therapy in equine musculoskeletal disease: scientific fact or clinical fiction?. Equine Vet J 2007 Mar;39(2):172-80.
    pubmed: 17378447doi: 10.2746/042516407x180868google scholar: lookup
  7. Radcliffe CH, Flaminio MJ, Fortier LA. Temporal analysis of equine bone marrow aspirate during establishment of putative mesenchymal progenitor cell populations.. Stem Cells Dev 2010 Feb;19(2):269-82.
    pmc: PMC3138180pubmed: 19604071doi: 10.1089/scd.2009.0091google scholar: lookup
  8. de Mattos Carvalho A, Alves AL, Golim MA, Moroz A, Hussni CA, de Oliveira PG, Deffune E. Isolation and immunophenotypic characterization of mesenchymal stem cells derived from equine species adipose tissue.. Vet Immunol Immunopathol 2009 Dec 15;132(2-4):303-6.
    pubmed: 19647331doi: 10.1016/j.vetimm.2009.06.014google scholar: lookup
  9. Stillwell C, Wang F, Xiang B, Deng J, Kashour T, Liu H, Freed D, Arora RC, Tian G. Adipose tissue houses different subtypes of stem cells.. Can J Physiol Pharmacol 2012 Sep;90(9):1295-301.
    pubmed: 22913488doi: 10.1139/y2012-094google scholar: lookup
  10. Ranera B, Lyahyai J, Romero A, Vázquez FJ, Remacha AR, Bernal ML, Zaragoza P, Rodellar C, Martín-Burriel I. Immunophenotype and gene expression profiles of cell surface markers of mesenchymal stem cells derived from equine bone marrow and adipose tissue.. Vet Immunol Immunopathol 2011 Nov 15;144(1-2):147-54.
    pubmed: 21782255doi: 10.1016/j.vetimm.2011.06.033google scholar: lookup
  11. Aquino JB, Bolontrade MF, García MG, Podhajcer OL, Mazzolini G. Mesenchymal stem cells as therapeutic tools and gene carriers in liver fibrosis and hepatocellular carcinoma.. Gene Ther 2010 Jun;17(6):692-708.
    pubmed: 20220785doi: 10.1038/gt.2010.10google scholar: lookup
  12. Sun JS, Wu SY, Lin FH. The role of muscle-derived stem cells in bone tissue engineering.. Biomaterials 2005 Jun;26(18):3953-60.
    pubmed: 15626442doi: 10.1016/j.biomaterials.2004.10.016google scholar: lookup
  13. Zhang L, Chan C. Isolation and enrichment of rat mesenchymal stem cells (MSCs) and separation of single-colony derived MSCs.. J Vis Exp 2010 Mar 22;(37).
    pmc: PMC3168201pubmed: 20308982doi: 10.3791/1852google scholar: lookup
  14. Jiang Y, Jahagirdar BN, Reinhardt RL, Schwartz RE, Keene CD, Ortiz-Gonzalez XR, Reyes M, Lenvik T, Lund T, Blackstad M, Du J, Aldrich S, Lisberg A, Low WC, Largaespada DA, Verfaillie CM. Pluripotency of mesenchymal stem cells derived from adult marrow.. Nature 2002 Jul 4;418(6893):41-9.
    pubmed: 12077603doi: 10.1038/nature00870google scholar: lookup
  15. Reschiglian P, Zattoni A, Roda B, Michelini E, Roda A. Field-flow fractionation and biotechnology.. Trends Biotechnol 2005 Sep;23(9):475-83.
    pubmed: 16061297doi: 10.1016/j.tibtech.2005.07.008google scholar: lookup
  16. Roda B, Reschiglian P, Zattoni A, Alviano F, Lanzoni G, Costa R, Di Carlo A, Marchionni C, Franchina M, Bonsi L, Bagnara GP. A tag-less method of sorting stem cells from clinical specimens and separating mesenchymal from epithelial progenitor cells.. Cytometry B Clin Cytom 2009 Jul;76(4):285-90.
    pubmed: 19363787doi: 10.1002/cyto.b.20472google scholar: lookup
  17. Roda B, Reschiglian P, Zattoni A, Tazzari PL, Buzzi M, Ricci F, Bontadini A. Human lymphocyte sorting by gravitational field-flow fractionation.. Anal Bioanal Chem 2008 Sep;392(1-2):137-45.
    pubmed: 18649074doi: 10.1007/s00216-008-2271-xgoogle scholar: lookup
  18. Roda B, Reschiglian P, Alviano F, Lanzoni G, Bagnara GP, Ricci F, Buzzi M, Tazzari PL, Pagliaro P, Michelini E, Roda A. Gravitational field-flow fractionation of human hemopoietic stem cells.. J Chromatogr A 2009 Dec 25;1216(52):9081-7.
    pubmed: 19647835doi: 10.1016/j.chroma.2009.07.024google scholar: lookup
  19. Vidal MA, Kilroy GE, Lopez MJ, Johnson JR, Moore RM, Gimble JM. Characterization of equine adipose tissue-derived stromal cells: adipogenic and osteogenic capacity and comparison with bone marrow-derived mesenchymal stromal cells.. Vet Surg 2007 Oct;36(7):613-22.
    pubmed: 17894587doi: 10.1111/j.1532-950x.2007.00313.xgoogle scholar: lookup
  20. Bhargava U, Bar-Lev M, Bellows CG, Aubin JE. Ultrastructural analysis of bone nodules formed in vitro by isolated fetal rat calvaria cells.. Bone 1988;9(3):155-63.
    pubmed: 3166832doi: 10.1016/8756-3282(88)90005-1google scholar: lookup
  21. BURSTONE MS. Histochemical observations on enzymatic processes in bones and teeth.. Ann N Y Acad Sci 1960 Mar 29;85:431-44.
    pubmed: 13806316doi: 10.1111/j.1749-6632.1960.tb49972.xgoogle scholar: lookup
  22. Owens SD, Burges J, Johns JL, Carrade DD, Galuppo LD, Librach F, Borjesson DL. Processing of equine bone marrow using the automated MarrowXpress System: RBC depletion, volume reduction, and mononuclear cell recovery.. Vet Clin Pathol 2011 Dec;40(4):444-449.
    pubmed: 22092275doi: 10.1111/j.1939-165x.2011.00368.xgoogle scholar: lookup
  23. Roda B, Cioffi N, Ditaranto N, Zattoni A, Casolari S, Melucci D, Reschiglian P, Sabbatini L, Valentini A, Zambonin PG. Biocompatible channels for field-flow fractionation of biological samples: correlation between surface composition and operating performance.. Anal Bioanal Chem 2005 Feb;381(3):639-46.
    pubmed: 15702313doi: 10.1007/s00216-004-2860-2google scholar: lookup
  24. Stewart AA, Barrett JG, Byron CR, Yates AC, Durgam SS, Evans RB, Stewart MC. Comparison of equine tendon-, muscle-, and bone marrow-derived cells cultured on tendon matrix.. Am J Vet Res 2009 Jun;70(6):750-7.
    pubmed: 19496665doi: 10.2460/ajvr.70.6.750google scholar: lookup
  25. Toupadakis CA, Wong A, Genetos DC, Cheung WK, Borjesson DL, Ferraro GL, Galuppo LD, Leach JK, Owens SD, Yellowley CE. Comparison of the osteogenic potential of equine mesenchymal stem cells from bone marrow, adipose tissue, umbilical cord blood, and umbilical cord tissue.. Am J Vet Res 2010 Oct;71(10):1237-45.
    pubmed: 20919913doi: 10.2460/ajvr.71.10.1237google scholar: lookup
  26. McD￮ LA, Anderson GI. In vitro comparison of equine cancellous bone graft donor sites and tibial periosteum as sources of viable osteoprogenitors.. Vet Surg 2003 Sep-Oct;32(5):455-63.
    pubmed: 14569574doi: 10.1053/jvet.2003.50060google scholar: lookup

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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