Chondrogenic potential of mesenchymal stromal cells derived from equine bone marrow and umbilical cord blood

L. C. Berg; T. G. Koch; T. Heerkens; K. Besonov; P. D. Thomsen; D. H. Betts

doi:10.3415/VCOT-08-10-0107

Veterinary and Comparative Orthopaedics and Traumatology, Table of Contents

Vet Comp Orthop Traumatol 2009; 22(05): 363-370
DOI: 10.3415/VCOT-08-10-0107

Original Research

Schattauer GmbH

Chondrogenic potential of mesenchymal stromal cells derived from equine bone marrow and umbilical cord blood

L. C. Berg

¹Department of Basic Animal and Veterinary Sciences, Faculty of Life Sciences, University of Copenhagen, Frederiksberg C, Denmark

,

T. G. Koch

¹Department of Basic Animal and Veterinary Sciences, Faculty of Life Sciences, University of Copenhagen, Frederiksberg C, Denmark

²Department of Biomedical Sciences, University of Guelph, Guelph, Canada

,

T. Heerkens

²Department of Biomedical Sciences, University of Guelph, Guelph, Canada

,

K. Besonov

²Department of Biomedical Sciences, University of Guelph, Guelph, Canada

,

P. D. Thomsen

¹Department of Basic Animal and Veterinary Sciences, Faculty of Life Sciences, University of Copenhagen, Frederiksberg C, Denmark

,

D. H. Betts

²Department of Biomedical Sciences, University of Guelph, Guelph, Canada

› Author Affiliations

Abstract

Summary

Objective: Orthopaedic injury is the most common cause of lost training days or premature retirement in the equine athlete. Cell-based therapies are a potential new treatment option in musculo-skeletal diseases. Mesenchymal stromal cells (MSC) have been derived from multiple sources in the horse including bone marrow and umbilical cord blood. The objective of this study was to provide an in vitro comparison of the chondrogenic potential in MSC derived from adult bone marrow (BM-MSC) and umbilical cord blood (CB-MSC).

Results: MSC from both sources produced tissue with cartilage-like morphology that stained positive for proteoglycans and expressed cartilage markers. The CB-MSC pellets were larger and showed hyaline-like cartilage morphology as early as day six. Gene expression of collagen type 21, aggrecan and CD-RAP was higher in CB- than BM-MSC pel-lets. Expression of Sox9 mRNA was similar between CB- and BM-MSC pellets. Protein concentration of cartilage-derived retinoic acid sensitive protein was higher in culture medium from CB- than BM-MSC pellets.

Conclusion: CB-MSC and BM-MSC were both capable of producing hyaline-like cartilage in vitro. However, in this study the MSC from umbilical cord blood appeared to have more chondrogenic potential than the BMMSC based on the cells tested and parameters measured.

Supplementary Information for this paper is available on the VCOT website at www.VCOTonline.com.

Keywords

Chondrogenesis - mesenchymal stromal cells (MSC) - bone marrow - cord blood - horse

Full Text

References

References
1 Jeffcott LB, Rossdale PD, Freestone J. et al. An assessment of wastage in thoroughbred racing from conception to 4 years of age. Equine Vet J 1982; Jul 14 (03) 185-198.
2 Rossdale PD, Hopes R, Digby NJ. et al. Epidemiological study of wastage among racehorses 1982 and 1983. Vet Rec 1985; Jan 116 (03) 66-69.
3 Bailey CJ, Reid SW, Hodgson DR. et al. Impact of in-juries and disease on a cohort of two- and three-year-old thoroughbreds in training. Vet Rec 1999; Oct 23 145 (17) 487-493.
4 Fortier LA, Smith RK. Regenerative medicine for tendinous and ligamentous injuries of sport horses. Vet Clin North Am Equine Pract 2008; Apr 24 (01) 191-201.
5 Pacini S, Spinabella S, Trombi L. et al. Suspension of bone marrow-derived undifferentiated mesenchymal stromal cells for repair of superficial digital flexor tendon in race horses. Tissue Eng 2007; Dec 13 (12) 2949-2955.
6 Smith RK, Korda M, Blunn GW. Isolation and implantation of autologous equine mesenchymal stem cells from bone marrow into the superficial digital flexor tendon as a potential novel treatment. Equine Vet J 2003; Jan 35 (01) 99-102.
7 Crovace A, Lacitignola L, De Siena R. et al. Cell therapy for tendon repair in horses: an experimental study. Vet Res Commun 2007; Aug 31 (Suppl. 01) Suppl 281-283.
8 Wilke MM, Nydam DV, Nixon AJ. Enhanced early chondrogenesis in articular defects following arthroscopic mesenchymal stem cell implantation in an equine model. J Orthop Res 2007; Jul 25 (07) 913-925.
9 Guest DJ, Smith MR, Allen WR. Monitoring the fate of autologous and allogeneic mesenchymal progenitor cells injected into the superficial digital flex-or tendon of horses: preliminary study. Equine Vet J 2008; Mar 40 (02) 178-181.
10 Smith JJ, Ross MW, Smith RK. Anabolic effects of acellular bone marrow, platelet rich plasma, and serum on equine suspensory ligament fibroblasts in vitro. Vet Comp Orthop Traumatol 2006; 19 (01) 43-47.
11 Fortier LA, Nixon AJ, Williams J. et al. Isolation and chondrocytic differentiation of equine bone marrow-derived mesenchymal stem cells. Am J Vet Res 1998; Sep 59 (09) 1182-1187.
12 Koerner J, Nesic D, Romero JD. et al. Equine peripheral blood-derived progenitors in comparison to bone marrow-derived mesenchymal stem cells. Stem Cells 2006; Jun 24 (06) 1613-1619.
13 Koch TG, Heerkens T, Thomsen PD. et al. Isolation of mesenchymal stem cells from equine umbilical cord blood. BMC Biotechnol 2007 May 7-26.
14 Giovannini S, Brehm W, Mainil-Varlet P. et al. Multilineage differentiation potential of equine blood-derived fibroblast-like cells. Differentiation 2008; Feb 76 (02) 118-129.
15 Kisiday JD, Kopesky PW, Evans CH. et al. Evaluation of adult equine bone marrow- and adipose-derived progenitor cell chondrogenesis in hydrogel cultures. J Orthop Res 2008; Mar 26 (03) 322-331.
16 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 (02) 329-336.
17 Kern S, Eichler H, Stoeve J. et al. Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue. Stem Cells 2006; May 24 (05) 1294-1301.
18 Stolzing A, Jones E, McGonagle D. et al. Age-related changes in human bone marrow-derived mesenchymal stem cells: consequences for cell therapies. Mech Ageing Dev 2008; Mar 129 (03) 163-173.
19 Sethe S, Scutt A, Stolzing A. Aging of mesenchymal stem cells. Ageing Res Rev 2006; Feb 5 (01) 91-116.
20 Bonyadi M, Waldman SD, Liu D. et al. Mesenchymal progenitor self-renewal deficiency leads to age-dependent osteoporosis in Sca-1/Ly-6A null mice. Proc Natl Acad Sci U S A 2003; May 100 (10) 5840-5845.
21 Shapiro F, Koide S, Glimcher MJ. Cell origin and differentiation in the repair of full-thickness defects of articular cartilage. J Bone Joint Surg Am 1993; Apr 75 (04) 532-553.
22 Steinert AF, Ghivizzani SC, Rethwilm A. et al. Major biological obstacles for persistent cell-based regeneration of articular cartilage. Arthritis Res Ther 2007; 9 (03) 213 15 pages
23 Kogler G, Sensken S, Wernet P. Comparative generation and characterization of pluripotent unrestricted somatic stem cells with mesenchymal stem cells from human cord blood. Exp Hematol 2006; Nov 34 (11) 1589-1595.
24 Kogler G, Sensken S, Airey JA. et al. A new human somatic stem cell from placental cord blood with intrinsic pluripotent differentiation potential. J Exp Med 2004; Jul 19 200 (02) 123-135.
25 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 (02) 329-336.
26 Bosserhoff AK, Buettner R. Establishing the protein MIA (melanoma inhibitory activity) as a marker for chondrocyte differentiation. Biomaterials 2003; Aug 24 (19) 3229-3234.
27 Berg LC, Mata X, Thomsen PD. Molecular characterization and chromosomal assignment of equine cartilage derived retinoic acid sensitive protein (CD-RAP)/melanoma inhibitory activity (MIA). Gene 2008; Jan 407 1–2 98-104.
28 Bieback K, Kern S, Kluter H. et al. Critical parameters for the isolation of mesenchymal stem cells from umbilical cord blood. Stem Cells 2004; 22 (04) 625-634.
29 Johnstone B, Hering TM, Caplan I A. et al. In vitro chondrogenesis of bone marrow-derived mesenchymal progenitor cells. Exp Cell Res 1998; Jan 238 (01) 265-272.
30 Stewart AA, Byron CR, Pondenis HC. et al. Effect of dexamethasone supplementation on chondrogenesis of equine mesenchymal stem cells. Am J Vet Res 2008; Aug 69 (08) 1013-1021.
31 Medrado GC, Machado CB, Valerio P. et al. The effect of a chitosan-gelatin matrix and dexamethasone on the behavior of rabbit mesenchymal stem cells. Biomed Mater 2006; Sep 1 (03) 155-161.
32 Derfoul A, Perkins GL, Hall DJ. et al. Glucocorticoids promote chondrogenic differentiation of adult human mesenchymal stem cells by enhancing expression of cartilage extracellular matrix genes. Stem Cells 2006; Jun 24 (06) 1487-1495.
33 Stewart AA, Byron CR, Pondenis H. et al. Effect of fibroblast growth factor-2 on equine mesenchymal stem cell monolayer expansion and chondrogenesis. Am J Vet Res 2007; Sep 68 (09) 941-945.
34 Dickhut A, Pelttari K, Janicki P. et al. Calcification or dedifferentiation: requirement to lock mesenchymal stem cells in a desired differentiation stage. J Cell Physiol 2009; Apr 219 (01) 219-226.
35 Murdoch AD, Hardingham TE. Clonal populations of human bone marrow mesenchymal cells with defined degrees of chondrogenic potential. Proceedings of the British Society for Matrix Biology 2007 Autumn Meeting P20-45.
36 Steenhuis P, Pettway GJ, Ignelzi Jr MA. Cell surface expression of stem cell antigen-1 (Sca-1) distinguishes osteo-, chondro-, and adipoprogenitors in fetal mouse calvaria. Calcif Tissue Int 2008; Jan 82 (01) 44-56.
37 Hachisuka H, Mochizuki Y, Yasunaga Y. et al. Flow cytometric discrimination of mesenchymal progenitor cells from bone marrow-adherent cell populations using CD34/44/45(-) and Sca-1(+) markers. J Orthop Sci 2007; Mar 12 (02) 161-169.
38 Jo CH, Ahn HJ, Kim HJ. et al. Surface characterization and chondrogenic differentiation of mesenchymal stromal cells derived from synovium. Cyto-therapy 2007; 9 (04) 316-327.
39 Varas L, Ohlsson LB, Honeth G. et al. Alpha10 inte-grin expression is up-regulated on fibroblast growth factor-2–treated mesenchymal stem cells with improved chondrogenic differentiation potential. Stem Cells Dev 2007; Dec 16 (06) 965-978.
40 Lim SM, Choi YS, Shin HC. et al. Isolation of human periosteum-derived progenitor cells using immunophenotypes for chondrogenesis. Biotechnol Lett 2005; May 27 (09) 607-611.
41 Rebelatto CK, Aguiar AM, Moretao MP. et al. Dissimilar differentiation of mesenchymal stem cells from bone marrow, umbilical cord blood, and adi-pose tissue. Exp Biol Med (Maywood ) 2008; Jul 233 (07) 901-913.

Supplementary Material

Online Supplementary Material