FREE SHIPPING over $40
OVER 9000 FIVE-STAR REVIEWS
5% OFF ALL SUBSCRIPTIONS
100% HAPPINESS GUARANTEE
Analyze Diet
0
Home / Equine Research Bank / Am J Vet Res / Isolation and chondrocytic differentiation of equine bone ma...
American journal of veterinary research1998; 59(9); 1182-1187;

Isolation and chondrocytic differentiation of equine bone marrow-derived mesenchymal stem cells.

Abstract: To isolate mesenchymal stem cells from adult horses and determine specific monolayer culture conditions required to enhance biochemically and phenotypically defined chondrocytic differentiation. Methods: 2 adult horse bone marrow donors without skeletal or hematologic abnormalities. Methods: Bone marrow was aspirated from the sternebra, and mesenchymal stem cells were isolated by centrifugation and cultured in monolayers. Subcultures were established in 24-well plates on day 13. Culture medium was harvested every 2 days, and culture of 12 of the 24 wells was terminated on day 6 and of the remaining wells on day 12. Medium proteoglycan content was determined for all samples, and proteoglycan monomeric size was determined for pooled samples from days 2-6 and 8-12. Total nucleated cell numbers were determined at culture termination on days 6 and 12. Histologic, histochemical, and collagen immunohistochemical analyses of multiwell chamber slides harvested on day 6 or 12 were performed. Results: Mesenchymal cells were an abundant cellular constituent of bone marrow aspirates, and separation of hematopoietic elements was achieved by centrifugation and delayed medium exchange. The remaining mesenchymal stem cells progressed from large, spindyloid, fibroblastic-appearing cells to a rounder shaped cell which formed colony plaques; isolated cells remained more spindyloid. Mesenchymal cell transformation toward a chondrocytic phenotype was verified by a shift in expression from collagen type I to type II, and an increase in quantity and molecular size of proteoglycans synthesized over time. Conclusions: Mesenchymal stem cells obtained from adult horses have the capacity to undergo chondrogenic differentiation in monolayer cultures and may provide a locally recruitable or transplantable autogenous cell source for articular cartilage repair.
Get Full Text
Publication Date: 1998-09-15 PubMed ID: 9736400
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
  • Research Support
  • Non-U.S. Gov't

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 article discusses the process of isolating mesenchymal stem cells from adult horses and cultivating them in specific conditions to promote chondrocytic (cartilage-forming) differentiation. The stem cells were obtained from the bone marrow of the horses and their transition into chondrocytic cells was verified through various tests and observations.

Methodology

  • The study involved two adult horse bone marrow donors that did not have any skeletal or hematologic abnormalities. The bone marrow was extracted from a structure called the sternebra.
  • The mesenchymal stem cells were isolated from the extracted bone marrow through a process of centrifugation. They were then cultured in single-layered (monolayer) cultures.
  • The subcultures were set up in plates of 24 wells on day 13. The culture medium was harvested in two day intervals, terminating half of the 24 wells on day 6 and the remaining on day 12.
  • The proteoglycan content from all samples was measured. Proteoglycan monomeric size was also determined from samples pooled from days 2-6 and 8-12. Total nucleated cell numbers were determined at the end of the culture on days 6 and 12.
  • Histological, histochemical, and immunohistochemical studies of collagen composition of the cells were also conducted on chamber slides harvested on day 6 or 12.

Results

  • The researchers found that plenty of mesenchymal cells were present in the bone marrow aspirates, and the unwanted hematopoietic elements were successfully separated by centrifugation and delayed medium exchange.
  • It was observed that the remaining mesenchymal stem cells went from large, spindle-shaped cells to rounder shaped cells that formed colony plaques. However, the isolated cells retained their spindyloid shape.
  • The transformation of mesenchymal cells towards a chondrocytic phenotype was confirmed by the shift in collagen type from I to II and the increase in the size and quantity of the proteoglycans synthesized over time.

Conclusions

  • The researchers concluded that mesenchymal stem cells obtained from adult horses possess the ability to undergo chondrogenic differentiation when cultured in monolayers. This suggests that these cells could potentially be used as a source of autologous cells for articular cartilage repair, which can be sourced locally or transplanted.

Cite This Article

APA
Fortier LA, Nixon AJ, Williams J, Cable CS. (1998). Isolation and chondrocytic differentiation of equine bone marrow-derived mesenchymal stem cells. Am J Vet Res, 59(9), 1182-1187.

Publication

American journal of veterinary research
ISSN: 0002-9645
NlmUniqueID: 0375011
Country: United States
Language: English
Volume: 59
Issue: 9
Pages: 1182-1187

Researcher Affiliations

Fortier, L A
  • Comparative Orthopaedics Laboratory, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
Nixon, A J
    Williams, J
      Cable, C S

        MeSH Terms

        • Animals
        • Bone Marrow Cells / cytology
        • Cell Differentiation
        • Cell Separation / veterinary
        • Cells, Cultured
        • Chondrocytes / cytology
        • Horses / anatomy & histology
        • Mesoderm / cytology
        • Microscopy, Phase-Contrast / veterinary
        • Stem Cells / cytology

        Citations

        This article has been cited 36 times.
        1. Voga M, Majdic G. Articular Cartilage Regeneration in Veterinary Medicine.. Adv Exp Med Biol 2022;1401:23-55.
          doi: 10.1007/5584_2022_717pubmed: 35733035google scholar: lookup
        2. Fülber J, Agreste FR, Seidel SRT, Sotelo EDP, Barbosa ÂP, Michelacci YM, Baccarin RYA. Chondrogenic potential of mesenchymal stem cells from horses using a magnetic 3D cell culture system.. World J Stem Cells 2021 Jun 26;13(6):645-658.
          doi: 10.4252/wjsc.v13.i6.645pubmed: 34249233google scholar: lookup
        3. Shah SS, Mithoefer K. Current Applications of Growth Factors for Knee Cartilage Repair and Osteoarthritis Treatment.. Curr Rev Musculoskelet Med 2020 Dec;13(6):641-650.
          doi: 10.1007/s12178-020-09664-6pubmed: 32710292google scholar: lookup
        4. Al Naem M, Bourebaba L, Kucharczyk K, Röcken M, Marycz K. Therapeutic mesenchymal stromal stem cells: Isolation, characterization and role in equine regenerative medicine and metabolic disorders.. Stem Cell Rev Rep 2020 Apr;16(2):301-322.
          doi: 10.1007/s12015-019-09932-0pubmed: 31797146google scholar: lookup
        5. Voga M, Drnovsek N, Novak S, Majdic G. Silk fibroin induces chondrogenic differentiation of canine adipose-derived multipotent mesenchymal stromal cells/mesenchymal stem cells.. J Tissue Eng 2019 Jan-Dec;10:2041731419835056.
          doi: 10.1177/2041731419835056pubmed: 30899447google scholar: lookup
        6. Bwalya EC, Wijekoon HS, Fang J, Kim S, Hosoya K, Okumura M. Independent chondrogenic potential of canine bone marrow-derived mesenchymal stem cells in monolayer expansion cultures decreases in a passage-dependent pattern.. J Vet Med Sci 2018 Nov 9;80(11):1681-1687.
          doi: 10.1292/jvms.18-0202pubmed: 30210068google scholar: lookup
        7. Barboni B, Russo V, Berardinelli P, Mauro A, Valbonetti L, Sanyal H, Canciello A, Greco L, Muttini A, Gatta V, Stuppia L, Mattioli M. Placental Stem Cells from Domestic Animals: Translational Potential and Clinical Relevance.. Cell Transplant 2018 Jan;27(1):93-116.
          doi: 10.1177/0963689717724797pubmed: 29562773google scholar: lookup
        8. Reesink HL, Sutton RM, Shurer CR, Peterson RP, Tan JS, Su J, Paszek MJ, Nixon AJ. Galectin-1 and galectin-3 expression in equine mesenchymal stromal cells (MSCs), synovial fibroblasts and chondrocytes, and the effect of inflammation on MSC motility.. Stem Cell Res Ther 2017 Nov 2;8(1):243.
          doi: 10.1186/s13287-017-0691-2pubmed: 29096716google scholar: lookup
        9. Zayed M, Caniglia C, Misk N, Dhar MS. Donor-Matched Comparison of Chondrogenic Potential of Equine Bone Marrow- and Synovial Fluid-Derived Mesenchymal Stem Cells: Implications for Cartilage Tissue Regeneration.. Front Vet Sci 2016;3:121.
          doi: 10.3389/fvets.2016.00121pubmed: 28149840google scholar: lookup
        10. Williams LB, Co C, Koenig JB, Tse C, Lindsay E, Koch TG. Response to Intravenous Allogeneic Equine Cord Blood-Derived Mesenchymal Stromal Cells Administered from Chilled or Frozen State in Serum and Protein-Free Media.. Front Vet Sci 2016;3:56.
          doi: 10.3389/fvets.2016.00056pubmed: 27500136google scholar: lookup
        11. Liebesny PH, Byun S, Hung HH, Pancoast JR, Mroszczyk KA, Young WT, Lee RT, Frisbie DD, Kisiday JD, Grodzinsky AJ. Growth Factor-Mediated Migration of Bone Marrow Progenitor Cells for Accelerated Scaffold Recruitment.. Tissue Eng Part A 2016 Jul;22(13-14):917-27.
          doi: 10.1089/ten.TEA.2015.0524pubmed: 27268956google scholar: lookup
        12. 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
        13. Grissom MJ, Temple-Wong MM, Adams MS, Tom M, Schumacher BL, McIlwraith CW, Goodrich LR, Chu CR, Sah RL. Synovial Fluid Lubricant Properties are Transiently Deficient after Arthroscopic Articular Cartilage Defect Repair with Platelet-Enriched Fibrin Alone and with Mesenchymal Stem Cells.. Orthop J Sports Med 2014 Jul;2(7).
          doi: 10.1177/2325967114542580pubmed: 25530978google scholar: lookup
        14. Rutigliano L, Corradetti B, Valentini L, Bizzaro D, Meucci A, Cremonesi F, Lange-Consiglio A. Molecular characterization and in vitro differentiation of feline progenitor-like amniotic epithelial cells.. Stem Cell Res Ther 2013 Oct 30;4(5):133.
          doi: 10.1186/scrt344pubmed: 24405576google 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.
          doi: 10.2147/sccaa.s3824pubmed: 24198500google scholar: lookup
        16. Smith RK, Werling NJ, Dakin SG, Alam R, Goodship AE, Dudhia J. Beneficial effects of autologous bone marrow-derived mesenchymal stem cells in naturally occurring tendinopathy.. PLoS One 2013;8(9):e75697.
          doi: 10.1371/journal.pone.0075697pubmed: 24086616google scholar: lookup
        17. Carrade DD, Borjesson DL. Immunomodulation by mesenchymal stem cells in veterinary species.. Comp Med 2013 Jun;63(3):207-17.
          pubmed: 23759523
        18. Watts AE, Ackerman-Yost JC, Nixon AJ. A comparison of three-dimensional culture systems to evaluate in vitro chondrogenesis of equine bone marrow-derived mesenchymal stem cells.. Tissue Eng Part A 2013 Oct;19(19-20):2275-83.
          doi: 10.1089/ten.TEA.2012.0479pubmed: 23725547google scholar: lookup
        19. Carrade DD, Lame MW, Kent MS, Clark KC, Walker NJ, Borjesson DL. Comparative Analysis of the Immunomodulatory Properties of Equine Adult-Derived Mesenchymal Stem Cells().. Cell Med 2012;4(1):1-11.
          doi: 10.3727/215517912X647217pubmed: 23152950google scholar: lookup
        20. Campbell KJ, Boykin RE, Wijdicks CA, Erik Giphart J, LaPrade RF, Philippon MJ. Treatment of a hip capsular injury in a professional soccer player with platelet-rich plasma and bone marrow aspirate concentrate therapy.. Knee Surg Sports Traumatol Arthrosc 2013 Jul;21(7):1684-8.
          doi: 10.1007/s00167-012-2232-ypubmed: 23052123google scholar: lookup
        21. Smyth NA, Murawski CD, Haleem AM, Hannon CP, Savage-Elliott I, Kennedy JG. Establishing proof of concept: Platelet-rich plasma and bone marrow aspirate concentrate may improve cartilage repair following surgical treatment for osteochondral lesions of the talus.. World J Orthop 2012 Jul 18;3(7):101-8.
          doi: 10.5312/wjo.v3.i7.101pubmed: 22816065google scholar: lookup
        22. Zhang Y, Fan Y, Wang Z, Wan Y, Zhou Z, Zhong B, Wang L, Wang F. Isolation, characterization, and gene modification of dairy goat mesenchymal stem cells from bone marrow.. In Vitro Cell Dev Biol Anim 2012 Aug;48(7):418-25.
          doi: 10.1007/s11626-012-9530-zpubmed: 22806970google scholar: lookup
        23. Delling U, Lindner K, Ribitsch I, Jülke H, Brehm W. Comparison of bone marrow aspiration at the sternum and the tuber coxae in middle-aged horses.. Can J Vet Res 2012 Jan;76(1):52-6.
          pubmed: 22754095
        24. Violini S, Gorni C, Pisani LF, Ramelli P, Caniatti M, Mariani P. Isolation and differentiation potential of an equine amnion-derived stromal cell line.. Cytotechnology 2012 Jan;64(1):1-7.
          doi: 10.1007/s10616-011-9398-xpubmed: 21994048google scholar: lookup
        25. Fortier LA, Barker JU, Strauss EJ, McCarrel TM, Cole BJ. The role of growth factors in cartilage repair.. Clin Orthop Relat Res 2011 Oct;469(10):2706-15.
          doi: 10.1007/s11999-011-1857-3pubmed: 21403984google scholar: lookup
        26. Lodi D, Iannitti T, Palmieri B. Stem cells in clinical practice: applications and warnings.. J Exp Clin Cancer Res 2011 Jan 17;30(1):9.
          doi: 10.1186/1756-9966-30-9pubmed: 21241480google scholar: lookup
        27. Raabe O, Reich C, Wenisch S, Hild A, Burg-Roderfeld M, Siebert HC, Arnhold S. Hydrolyzed fish collagen induced chondrogenic differentiation of equine adipose tissue-derived stromal cells.. Histochem Cell Biol 2010 Dec;134(6):545-54.
          doi: 10.1007/s00418-010-0760-4pubmed: 21076963google scholar: lookup
        28. Colleoni S, Bottani E, Tessaro I, Mari G, Merlo B, Romagnoli N, Spadari A, Galli C, Lazzari G. Isolation, growth and differentiation of equine mesenchymal stem cells: effect of donor, source, amount of tissue and supplementation with basic fibroblast growth factor.. Vet Res Commun 2009 Dec;33(8):811-21.
          doi: 10.1007/s11259-009-9229-0pubmed: 19472068google scholar: lookup
        29. Koch TG, Berg LC, Betts DH. Current and future regenerative medicine - principles, concepts, and therapeutic use of stem cell therapy and tissue engineering in equine medicine.. Can Vet J 2009 Feb;50(2):155-65.
          pubmed: 19412395
        30. 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: 10.1186/1471-2121-10-29pubmed: 19383177google scholar: lookup
        31. Vidal MA, Robinson SO, Lopez MJ, Paulsen DB, Borkhsenious O, Johnson JR, Moore RM, Gimble JM. Comparison of chondrogenic potential in equine mesenchymal stromal cells derived from adipose tissue and bone marrow.. Vet Surg 2008 Dec;37(8):713-24.
          doi: 10.1111/j.1532-950X.2008.00462.xpubmed: 19121166google scholar: lookup
        32. Koch TG, Berg LC, Betts DH. Concepts for the clinical use of stem cells in equine medicine.. Can Vet J 2008 Oct;49(10):1009-17.
          pubmed: 19119371
        33. Csaki C, Matis U, Mobasheri A, Ye H, Shakibaei M. Chondrogenesis, osteogenesis and adipogenesis of canine mesenchymal stem cells: a biochemical, morphological and ultrastructural study.. Histochem Cell Biol 2007 Dec;128(6):507-20.
          doi: 10.1007/s00418-007-0337-zpubmed: 17922135google scholar: lookup
        34. 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: 10.1186/1472-6750-7-26pubmed: 17537254google scholar: lookup
        35. Carstanjen B, Desbois C, Hekmati M, Behr L. Successful engraftment of cultured autologous mesenchymal stem cells in a surgically repaired soft palate defect in an adult horse.. Can J Vet Res 2006 Apr;70(2):143-7.
          pubmed: 16639947
        36. Lin Y, Luo E, Chen X, Liu L, Qiao J, Yan Z, Li Z, Tang W, Zheng X, Tian W. Molecular and cellular characterization during chondrogenic differentiation of adipose tissue-derived stromal cells in vitro and cartilage formation in vivo.. J Cell Mol Med 2005 Oct-Dec;9(4):929-39.
          doi: 10.1111/j.1582-4934.2005.tb00389.xpubmed: 16364200google scholar: lookup
        Subscribe to our newsletter
        Join 99,344 horse owners like you who receive our email updates on horse health and nutrition.