FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Int J Stem Cells / Stemness Signature of Equine Marrow-derived Mesenchymal Stem...
International journal of stem cells2017; 10(1); 93-102; doi: 10.15283/ijsc16036

Stemness Signature of Equine Marrow-derived Mesenchymal Stem Cells.

Abstract: Application of competent cells such as mesenchymal stem cells (MSCs) for treatment of musculoskeletal disorders in equine athletes is increasingly needed. Moreover, similarities of horse and human in size, load and types of joint injuries, make horse as a good model for MSCs therapy studies. This study was designed to isolate and characterize stemness signature of equine bone marrow-derived mesenchymal stem cells (BM-MSCs). Methods: BM of three mares was aspirated and the mononuclear cells (MNCs) were isolated using density gradient. The primary MNCs were cultured and analyzed after tree passages (P3) for growth characteristics, differentiation potentials, and the expression of genes including CD29, CD34, CD44, CD90, CD105, MHC-I, MHC-II and pluripotency related genes (Nanog, Oct-4, Sox-2, SSEA-1, -3, -4) using RT-PCR or immunocytochemistry techniques. Results: The isolated cells in P3 were adherent and fibroblast-like in shape with doubling times of 78.15 h. Their clonogenic capacity was 8.67±4% and they were able to differentiate to osteogenic, adipogenic and chondrogenic lineages. Cells showed expression of CD29, CD44, CD90, MHC-I and Sox-2 while no expression for CD34, MHC-II, CD105, and pluripotency stemness markers was detected. Conclusions: In conclusion, data showed that isolated cells have the basic and minimal criteria for MSCs, however, expressing only one pluripotency gene (sox-2).
Get Full Text
Publication Date: 2017-02-22 PubMed ID: 28222255PubMed Central: PMC5488781DOI: 10.15283/ijsc16036Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
LinkedInCopy
  • 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.

This research conducted an examination of bone marrow-derived mesenchymal stem cells (BM-MSCs) from horses to understand their growth characteristics, gene expression, and ability to differentiate into various cell types. While the harvested cells fulfilled basic criteria for MSCs, they only expressed one gene associated with pluripotency, Sox-2.

Studying Mesenchymal Stem Cells in Horses

  • The goal of the research was to explore the properties of mesenchymal stem cells (MSCs), derived from horse bone marrow, due to their potential in treating musculoskeletal disorders in equine athletes. Moreover, because of similarities between horses and humans in terms of size, joint injuries and load, horses can be effective models for studies about MSCs therapies.

Methods and Observations

  • Bone marrow was aspirated from three mares and mononuclear cells (MNCs) were isolated. These primary MNCs were cultured and assessed after three passages for their growth characteristics, differentiation potentials, and gene expression.
  • The results showed that cells isolated in the third passage were adherent, had a fiber-like shape with doubling times of 78.15 h, clonogenic capacity of 8.67±4%, and were capable of differentiating into osteogenic, adipogenic and chondrogenic lineages.

Expression of Various Genes

  • The evaluation was performed for several genes including CD29, CD34, CD44, CD90, CD105, MHC-I, MHC-II and pluripotency related genes (Nanog, Oct-4, Sox-2, SSEA-1, -3, -4) using RT-PCR or immunocytochemistry techniques.
  • Cells showed expression for CD29, CD44, CD90, MHC-I and Sox-2. On the contrary, the cells did not express CD34, MHC-II, CD105, and most pluripotency stemness markers.

Conclusions

  • The conclusion of the study was that while the isolated cells fulfilled the basic and minimal criteria for MSCs, the expression was limited to only one pluripotency-associated gene, Sox-2.

Cite This Article

APA
Zahedi M, Parham A, Dehghani H, Mehrjerdi HK. (2017). Stemness Signature of Equine Marrow-derived Mesenchymal Stem Cells. Int J Stem Cells, 10(1), 93-102. https://doi.org/10.15283/ijsc16036

Publication

International journal of stem cells
ISSN: 2005-3606
NlmUniqueID: 101497587
Country: Korea (South)
Language: English
Volume: 10
Issue: 1
Pages: 93-102

Researcher Affiliations

Zahedi, Morteza
  • Division of Physiology, Department of Basic Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran.
Parham, Abbas
  • Division of Physiology, Department of Basic Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran.
  • Embryonic and Stem Cell Biology and Biotechnology Research Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran.
Dehghani, Hesam
  • Division of Physiology, Department of Basic Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran.
  • Embryonic and Stem Cell Biology and Biotechnology Research Group, Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran.
Mehrjerdi, Hossein Kazemi
  • Department of Clinical Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran.

Conflict of Interest Statement

. The authors have no conflicting financial interest.

References

This article includes 40 references
  1. Metsäranta M, Kujala UM, Pelliniemi L, Osterman H, Aho H, Vuorio E. Evidence for insufficient chondrocytic differentiation during repair of full-thickness defects of articular cartilage.. Matrix Biol 1996 Apr;15(1):39-47.
    doi: 10.1016/S0945-053X(96)90125-0pubmed: 8783186google scholar: lookup
  2. Paris DB, Stout TA. Equine embryos and embryonic stem cells: defining reliable markers of pluripotency.. Theriogenology 2010 Sep 1;74(4):516-24.
    doi: 10.1016/j.theriogenology.2009.11.020pubmed: 20071015google scholar: lookup
  3. 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.
    doi: 10.1016/j.theriogenology.2010.11.008pubmed: 21196039google scholar: lookup
  4. 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.
    doi: 10.2746/042516407X180868pubmed: 17378447google scholar: lookup
  5. Frisbie DD, Kawcak CE, Werpy NM, McIlwraith CW. Evaluation of bone marrow derived stem cells and adipose derived stromal vascular fraction for treatment of osteoarthitis using an equine experimental model. AAEP Proceedings 2006. pp. 420–421.
  6. Koch TG, Betts DH. Stem cell therapy for joint problems using the horse as a clinically relevant animal model.. Expert Opin Biol Ther 2007 Nov;7(11):1621-6.
    doi: 10.1517/14712598.7.11.1621pubmed: 17961087google scholar: lookup
  7. Ahern BJ, Parvizi J, Boston R, Schaer TP. Preclinical animal models in single site cartilage defect testing: a systematic review.. Osteoarthritis Cartilage 2009 Jun;17(6):705-13.
    doi: 10.1016/j.joca.2008.11.008pubmed: 19101179google scholar: lookup
  8. Smith RK, Webbon PM. Harnessing the stem cell for the treatment of tendon injuries: heralding a new dawn?. Br J Sports Med 2005 Sep;39(9):582-4.
    doi: 10.1136/bjsm.2005.015834pmc: PMC1725307pubmed: 16118291google scholar: lookup
  9. Ibrahim S, Saunders K, Kydd JH, Lunn DP, Steinbach F. Screening of anti-human leukocyte monoclonal antibodies for reactivity with equine leukocytes.. Vet Immunol Immunopathol 2007 Sep 15;119(1-2):63-80.
    doi: 10.1016/j.vetimm.2007.06.034pubmed: 17707518google scholar: lookup
  10. CTGTA Committee. Cellular products for joint surface repair-Briefing document. 2005.
  11. Ribitsch I, Burk J, Delling U, Geißler C, Gittel C, Jülke H, Brehm W. Basic science and clinical application of stem cells in veterinary medicine.. Adv Biochem Eng Biotechnol 2010;123:219-63.
    pubmed: 20309674doi: 10.1007/10_2010_66google scholar: lookup
  12. Lakshmipathy U, Verfaillie C. Stem cell plasticity.. Blood Rev 2005 Jan;19(1):29-38.
    doi: 10.1016/j.blre.2004.03.001pubmed: 15572215google scholar: lookup
  13. Ryan JM, Barry FP, Murphy JM, Mahon BP. Mesenchymal stem cells avoid allogeneic rejection.. J Inflamm (Lond) 2005 Jul 26;2:8.
    doi: 10.1186/1476-9255-2-8pmc: PMC1215510pubmed: 16045800google scholar: lookup
  14. Guest DJ, Smith MR, Allen WR. Monitoring the fate of autologous and allogeneic mesenchymal progenitor cells injected into the superficial digital flexor tendon of horses: preliminary study.. Equine Vet J 2008 Mar;40(2):178-81.
    doi: 10.2746/042516408X276942pubmed: 18267891google scholar: lookup
  15. Guest DJ, Ousey JC, Smith MR. Defining the expression of marker genes in equine mesenchymal stromal cells.. Stem Cells Cloning 2008;1:1-9.
    pmc: PMC3781685pubmed: 24198500doi: 10.2147/sccaa.s3824google scholar: lookup
  16. Horwitz EM, Le Blanc K, Dominici M, Mueller I, Slaper-Cortenbach I, Marini FC, Deans RJ, Krause DS, Keating A. Clarification of the nomenclature for MSC: The International Society for Cellular Therapy position statement.. Cytotherapy 2005;7(5):393-5.
    doi: 10.1080/14653240500319234pubmed: 16236628google scholar: lookup
  17. Hoynowski SM, Fry MM, Gardner BM, Leming MT, Tucker JR, Black L, Sand T, Mitchell KE. Characterization and differentiation of equine umbilical cord-derived matrix cells.. Biochem Biophys Res Commun 2007 Oct 19;362(2):347-53.
    doi: 10.1016/j.bbrc.2007.07.182pubmed: 17719011google scholar: lookup
  18. Li X, Zhou SG, Imreh MP, Ahrlund-Richter L, Allen WR. Horse embryonic stem cell lines from the proliferation of inner cell mass cells.. Stem Cells Dev 2006 Aug;15(4):523-31.
    doi: 10.1089/scd.2006.15.523pubmed: 16978056google scholar: lookup
  19. Reed SA, Johnson SE. Equine umbilical cord blood contains a population of stem cells that express Oct4 and differentiate into mesodermal and endodermal cell types.. J Cell Physiol 2008 May;215(2):329-36.
    doi: 10.1002/jcp.21312pubmed: 17929245google scholar: lookup
  20. Reed SA, Johnson SE. Refinement of culture conditions for maintenance of undifferentiated equine umbilical cord blood stem cells. J Equine Vet Sci 2012;32:360–366.
    doi: 10.1016/j.jevs.2011.12.004google scholar: lookup
  21. Lange C, Bassler P, Lioznov MV, Bruns H, Kluth D, Zander AR, Fiegel HC. Liver-specific gene expression in mesenchymal stem cells is induced by liver cells.. World J Gastroenterol 2005 Aug 7;11(29):4497-504.
    doi: 10.3748/wjg.v11.i29.4497pmc: PMC4398698pubmed: 16052678google scholar: lookup
  22. Yoon YS, Wecker A, Heyd L, Park JS, Tkebuchava T, Kusano K, Hanley A, Scadova H, Qin G, Cha DH, Johnson KL, Aikawa R, Asahara T, Losordo DW. Clonally expanded novel multipotent stem cells from human bone marrow regenerate myocardium after myocardial infarction.. J Clin Invest 2005 Feb;115(2):326-38.
    doi: 10.1172/JCI200522326pmc: PMC546424pubmed: 15690083google scholar: lookup
  23. Vidal MA, Kilroy GE, Johnson JR, Lopez MJ, Moore RM, Gimble JM. Cell growth characteristics and differentiation frequency of adherent equine bone marrow-derived mesenchymal stromal cells: adipogenic and osteogenic capacity.. Vet Surg 2006 Oct;35(7):601-10.
    doi: 10.1111/j.1532-950X.2006.00197.xpubmed: 17026544google scholar: lookup
  24. Baghaban Eslaminejad MR, Taghiyar L, Dehghan MM, Falahi F, Kazemi Mehrjerdi H. Equine marrow-derived mesenchymal stem cells: isolation, differentiation and culture optimization. Iran J Vet Res 2009;10:1–11.
  25. Lange-Consiglio A, Corradetti B, Meucci A, Perego R, Bizzaro D, Cremonesi F. Characteristics of equine mesenchymal stem cells derived from amnion and bone marrow: in vitro proliferative and multilineage potential assessment.. Equine Vet J 2013 Nov;45(6):737-44.
    doi: 10.1111/evj.12052pubmed: 23527626google scholar: lookup
  26. Alipour F, Parham A, Kazemi Mehrjerdi H, Dehghani H. Equine adipose-derived mesenchymal stem cells: phenotype and growth characteristics, gene expression profile and differentiation potentials.. Cell J 2015 Winter;16(4):456-65.
    pmc: PMC4297484pubmed: 25685736doi: 10.22074/cellj.2015.491google scholar: lookup
  27. Prockop DJ. Marrow stromal cells as stem cells for nonhematopoietic tissues.. Science 1997 Apr 4;276(5309):71-4.
    doi: 10.1126/science.276.5309.71pubmed: 9082988google scholar: lookup
  28. Mensing N, Gasse H, Hambruch N, Haeger JD, Pfarrer C, Staszyk C. Isolation and characterization of multipotent mesenchymal stromal cells from the gingiva and the periodontal ligament of the horse.. BMC Vet Res 2011 Aug 2;7:42.
    doi: 10.1186/1746-6148-7-42pmc: PMC3161857pubmed: 21810270google scholar: lookup
  29. Lovati AB, Corradetti B, Lange Consiglio A, Recordati C, Bonacina E, Bizzaro D, Cremonesi F. Comparison of equine bone marrow-, umbilical cord matrix and amniotic fluid-derived progenitor cells.. Vet Res Commun 2011 Feb;35(2):103-21.
    doi: 10.1007/s11259-010-9457-3pubmed: 21193959google scholar: lookup
  30. Bourzac C, Smith LC, Vincent P, Beauchamp G, Lavoie JP, Laverty S. Isolation of equine bone marrow-derived mesenchymal stem cells: a comparison between three protocols.. Equine Vet J 2010 Sep;42(6):519-27.
    doi: 10.1111/j.2042-3306.2010.00098.xpubmed: 20716192google scholar: lookup
  31. Baghaban Eslaminejad M, Mardpour S, Ebrahimi M. Growth kinetics and in vitro aging of mesenchymal stem cells isolated from rat adipose versus bone marrow tissues. Iran J Vet Surg 2008;3:9–20.
  32. Burk J, Ribitsch I, Gittel C, Juelke H, Kasper C, Staszyk C, Brehm W. Growth and differentiation characteristics of equine mesenchymal stromal cells derived from different sources.. Vet J 2013 Jan;195(1):98-106.
    doi: 10.1016/j.tvjl.2012.06.004pubmed: 22841420google scholar: lookup
  33. Ranera B, Ordovás L, Lyahyai J, Bernal ML, Fernandes F, Remacha AR, Romero A, Vázquez FJ, Osta R, Cons C, Varona L, Zaragoza P, Martín-Burriel I, Rodellar C. Comparative study of equine bone marrow and adipose tissue-derived mesenchymal stromal cells.. Equine Vet J 2012 Jan;44(1):33-42.
    doi: 10.1111/j.2042-3306.2010.00353.xpubmed: 21668489google scholar: lookup
  34. Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, Prockop Dj, Horwitz E. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement.. Cytotherapy 2006;8(4):315-7.
    doi: 10.1080/14653240600855905pubmed: 16923606google scholar: lookup
  35. Koch TG, Heerkens T, Thomsen PD, Betts DH. Isolation of mesenchymal stem cells from equine umbilical cord blood.. BMC Biotechnol 2007 May 30;7:26.
    doi: 10.1186/1472-6750-7-26pmc: PMC1904213pubmed: 17537254google scholar: lookup
  36. 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.
    doi: 10.1089/scd.2009.0091pmc: PMC3138180pubmed: 19604071google scholar: lookup
  37. Masui S, Nakatake Y, Toyooka Y, Shimosato D, Yagi R, Takahashi K, Okochi H, Okuda A, Matoba R, Sharov AA, Ko MS, Niwa H. Pluripotency governed by Sox2 via regulation of Oct3/4 expression in mouse embryonic stem cells.. Nat Cell Biol 2007 Jun;9(6):625-35.
    doi: 10.1038/ncb1589pubmed: 17515932google scholar: lookup
  38. Ranera B, Remacha AR, Álvarez-Arguedas S, Romero A, Vázquez FJ, Zaragoza P, Martín-Burriel I, Rodellar C. Effect of hypoxia on equine mesenchymal stem cells derived from bone marrow and adipose tissue.. BMC Vet Res 2012 Aug 22;8:142.
    doi: 10.1186/1746-6148-8-142pmc: PMC3483288pubmed: 22913590google scholar: lookup
  39. Violini S, Ramelli P, Pisani LF, Gorni C, Mariani P. Horse bone marrow mesenchymal stem cells express embryo stem cell markers and show the ability for tenogenic differentiation by in vitro exposure to BMP-12.. BMC Cell Biol 2009 Apr 22;10:29.
    doi: 10.1186/1471-2121-10-29pmc: PMC2678092pubmed: 19383177google scholar: lookup
  40. Marfe G, Massaro-Giordano M, Ranalli M, Cozzoli E, Di Stefano C, Malafoglia V, Polettini M, Gambacurta A. Blood derived stem cells: an ameliorative therapy in veterinary ophthalmology.. J Cell Physiol 2012 Mar;227(3):1250-6.
    doi: 10.1002/jcp.22953pubmed: 21792938google scholar: lookup

Citations

This article has been cited 8 times.
  1. Stage HJ, Trappe S, Söllig K, Trachsel DS, Kirsch K, Zieger C, Merle R, Aschenbach JR, Gehlen H. Multilineage Differentiation Potential of Equine Adipose-Derived Stromal/Stem Cells from Different Sources.. Animals (Basel) 2023 Apr 15;13(8).
    doi: 10.3390/ani13081352pubmed: 37106915google scholar: lookup
  2. Park A, Oh HJ, Ji K, Choi EM, Kim D, Kim E, Kim MK. Effect of Passage Number of Conditioned Medium Collected from Equine Amniotic Fluid Mesenchymal Stem Cells: Porcine Oocyte Maturation and Embryo Development.. Int J Mol Sci 2022 Jun 12;23(12).
    doi: 10.3390/ijms23126569pubmed: 35743012google scholar: lookup
  3. Wang M, Dai T, Meng Q, Wang W, Li S. Regulatory effects of miR-28 on osteogenic differentiation of human bone marrow mesenchymal stem cells.. Bioengineered 2022 Jan;13(1):684-696.
    doi: 10.1080/21655979.2021.2012618pubmed: 34978269google scholar: lookup
  4. Nino-Fong R, Esparza Gonzalez BP, Rodriguez-Lecompte JC, Montelpare W, McD￮ L. Development of a biologically immortalized equine stem cell line.. Can J Vet Res 2021 Oct;85(4):293-301.
    pubmed: 34602734
  5. Wright A, Arthaud-Day ML, Weiss ML. Therapeutic Use of Mesenchymal Stromal Cells: The Need for Inclusive Characterization Guidelines to Accommodate All Tissue Sources and Species.. Front Cell Dev Biol 2021;9:632717.
    doi: 10.3389/fcell.2021.632717pubmed: 33665190google scholar: lookup
  6. Salami F, Ghodrati M, Parham A, Mehrzad J. The effect of equine bone marrow-derived mesenchymal stem cells on the expression of apoptotic genes in neutrophils.. Vet Med Sci 2021 May;7(3):626-633.
    doi: 10.1002/vms3.427pubmed: 33471967google scholar: lookup
  7. Kamm JL, Parlane NA, Riley CB, Gee EK, Dittmer KE, McIlwraith CW. Blood type and breed-associated differences in cell marker expression on equine bone marrow-derived mesenchymal stem cells including major histocompatibility complex class II antigen expression.. PLoS One 2019;14(11):e0225161.
    doi: 10.1371/journal.pone.0225161pubmed: 31747418google scholar: lookup
  8. Shojaee A, Parham A. Strategies of tenogenic differentiation of equine stem cells for tendon repair: current status and challenges.. Stem Cell Res Ther 2019 Jun 18;10(1):181.
    doi: 10.1186/s13287-019-1291-0pubmed: 31215490google scholar: lookup
Subscribe to our newsletter
Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.