The aim of this study was to determine whether synovial fluid (SF) of dogs contains cells that have characteristics of MSCs and to describe their differentiation potential. SF adherent cells from 5 young German shepherd dogs (average 3.8 ± 0.9 years) were expanded (37°C, 5% CO2, humidified atmosphere) three weeks before their phenotype was characterized by flow-cytometry for the presence of CD90 and CD34. Population doubling time (PDT), number of CFU-F and adipogenic, osteogenic and chondrogenic potentials have been determined in vitro. In early passages PTD was 31 ± 10 hours and expansion fold after 3 sub cultivations (9 days) theoretically could be 372 ± 134. At P1, 0.55 ± 0.05% of SF cells had the ability to form CFU-F. Sixty-six percent of cells expressed CD90 and none of the cells expressed markers of hematopoietic cells. Oil Red O staining has shown accumulation of fat droplets in cells grown in adipogenic medium, while deposits of calcium in the osteogenic medium were evidenced with Alizarin red staining. SF cultured in hondrogenic and control medium in three-dimensional conditions formed a cartilage-like tissue. Alcian blue staining of pellets’ slides have shown a significant amount of glycosaminoglycans (GAGs) and immunohistochemistry analysis documented collagen type II expression. The amount of GAGs in pellets grown in both conditions showed no difference. SF cells in vitro exhibited osteogenic, adipogenic and chondrogenic differentiation potentials, suggesting the presence of different mesenchymal progenitors. These results also demonstrated that SF cells have a spontaneous chondrogenic potential that should be further explored for possible tissue engineering protocols.
If the inline PDF is not rendering correctly, you can download the PDF file here.
1. Smith MD: The normal synovium. Open Rheumatol J 2011 5:100-6.
2. Hunziker EB Rosenberg LC: Repair of partial-thickness defects in articular cartilage: cell recruitment from the synovial membrane. J Bone Joint Surg Am 1996 78(5):721-33.
3. Hunziker EB Kapfinger E: Removal of proteoglycans from the surface of defects in articular cartilage transiently enhances coverage by repair cells. J Bone Joint Surg Br 1998 80(1):144-50.
4. Hatsushika D Muneta T Nakamura T Horie M Koga H Nakagawa Y Tsuji K Hishikawa S Kobayashi E Sekiya I: Repetitive allogeneic intraarticular injections of synovial mesenchymal stem cells promote meniscus regeneration in a porcine massive meniscus defect model. Osteoarthritis Cartilage 2014 22(7):941-50.
5. Horie M Driscoll MD Sampson HW Sekiya I Caroom CT Prockop DJ Thomas DB: Implantation of allogenic synovial stem cells promotes meniscal regeneration in a rabbit meniscal defect model. J Bone Joint Surg Am 2012 94(8):701-12.
6. Horie M Sekiya I Muneta T Ichinose S Matsumoto K Saito H Murakami T Kobayashi E: Intra-articular Injected synovial stem cells differentiate into meniscal cells directly and promote meniscal regeneration without mobilization to distant organs in rat massive meniscal defect. Stem Cells 2009 27(4):878-87.
7. Furuzawa-Carballeda J Macip-Rodríguez PM Cabral AR: Osteoarthritis and rheumatoid arthritis pannus have similar qualitative metabolic characteristics and pro-inflammatory cytokine response. Clin Exp Rheumatol 2008 26(4):554-60.
8. Karystinou A Dell'Accio F Kurth TB Wackerhage H Khan IM Archer CW Jones EA Mitsiadis TA De Bari C: Distinct mesenchymal progenitor cell subsets in the adult human synovium. Rheumatology (Oxford) 2009 48(9):1057-64.
9. Nimura A Muneta T Koga H Mochizuki T Suzuki K Makino H Umezawa A Sekiya I: Increased proliferation of human synovial mesenchymal stem cells with autologous human serum: comparisons with bone marrow mesenchymal stem cells and with fetal bovine serum. Arthritis Rheum 2008;58(2):501-10.
10. Harvanová D1 Tóthová T Sarišský M Amrichová J Rosocha J: Isolation and characterization of synovial mesenchymal stem cells. Folia Biol (Praha) 2011;57(3):119-24.
11. Ando W Kutcher JJ Krawetz R Sen A Nakamura N Frank CB Hart DA: Clonal analysis of synovial fluid stem cells to characterize and identify stable mesenchymal stromal cell/mesenchymal progenitor cell phenotypes in a porcine model: a cell source with enhanced commitment to the chondrogenic lineage. Cytotherapy 2014 16(6):776-88.
12. Morito T Muneta T Hara K Ju YJ Mochizuki T Makino H Umezawa A Sekiya I: Synovial fluid-derived mesenchymal stem cells increase after intra-articular ligament injury in humans. Rheumatology (Oxford) 2008 47(8):1137-43.
13. Berg RI Sykes JE Kass PH Vernau W: Effect of repeated arthrocentesis on cytologic analysis of synovial fluid in dogs. J Vet Intern Med 2009 23(4):814-7.
14. Brittberg M Lindahl A Nilsson A Ohlsson C Isaksson O Peterson L: Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation. N Engl J Med 1994 331(14):889-95.
15. Cook JL Kuroki K Visco D Pelletier JP Schulz L Lafeber FP: The OARSI histopathology initiative - recommendations for histological assessments of osteoarthritis in the dog. Osteoarthritis Cartilage 2010 18:S66-79.
16. Portron S Merceron C Gauthier O Lesoeur J Sourice S Masson M Fellah BH Geffroy O Lallemand E Weiss P Guicheux J Vinatier C: Effects of in vitro low oxygen tension preconditioning of adipose stromal cells on their in vivo chondrogenic potential: application in cartilage tissue repair. PLoS One 2013 8(4):e62368.
17. Krawetz RJ Wu YE Martin L Rattner JB Matyas JR Hart DA: Synovial fluid progenitors expressing CD90+ from normal but not osteoarthritic joints undergo chondrogenic differentiation without micro-mass culture. PLoS One 20127(8):e43616.
18. 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 8:315-7.
19. Bianco P Robey P Saggio I Riminucci M: Mesenchymal stem cells in human bone marrow (skeletal stem cells): a critical discussion of their nature identity and significance in incurable skeletal disease. Human Gene Therapy 2010 21:1057-1066.
20. Kovačević Filipović M: The Blood is Rich in Different Types of Mesoderm Derived Stem and Progenitor Cells. Acta veterinaria(Beograd) 2014 64:156-178.
21. Neidhart M Seemayer CA Hummel KM Michel BA Gay RE Gay S: Functional characterization of adherent synovial fluid cells in rheumatoid arthritis: destructive potential in vitro and in vivo. Arthritis Rheum 2003;48(7):1873-80.
22. Debeljak Martačić J Francuski J Lužajić T Vuković N Mojsilović S Drndarević N Petakov M Glibetić M Marković D Radovanović A Todorović V Kovačević Filipović M: Characterization of deciduous teeth stem cells isolated from crown dental pulp. Vojnosanit Pregl 2014 71:1-10.
23. Ivanović Z Bartolozzi B Bernabei PA Cipolleschi MG Milenkovic P Praloran V DelloSbarba P: A simple one-step clonal assay allows the sequential detection of committed (CFU-GM-like) progenitors and several subsets of primitive (HPP-CFC) murine progenitors. Stem Cells 199917(4):219-25.
24. Martinello T Bronzini I Maccatrozzo L Mollo A Sampaolesi M Mascarello F Decaminada M Patruno M: Canine adipose-derived-mesenchymal stem cells do not lose stem features after a long-term cryopreservation. Res Vet Sci 2011 91:18-24.
25. Muraglia A Cancedda R Quarto R: Clonal mesenchymal progenitors from human bone marrow differentiate in vitro according to a hierarchical model. J Cell Sci 2000 113 (Pt 7):1161-6.
26. Brittberg M Lindahl A Nilsson A Ohlsson C Isaksson O Peterson L: Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation. N Engl J Med. 1994 331:889-95.
27. Horňák S Harvanová D Ledecký V Hluchý M Valenčáková-Agyagosová A Amrichová J Rosocha J Vaško G Švihla R Petrovič V: Reparation of chondral defects in rabbits by autologous and allogenous chondrocytes seeded on collagen/hyaluronan scaffold or suspended in fibrin glue. Acta veterinaria (Beograd) 2014 64:61-72.
28. Jones EA Crawford A English A Henshaw K Mundy J Corscadden D Chapman T Emery P Hatton P McGonagle D: Synovial fluid mesenchymal stem cells in health and early osteoarthritis: detection and functional evaluation at the single-cell level. Arthritis Rheumatology 2008 58:1731-1740.
29. Ichinose S Muneta T Koga H Segawa Y Tagami M Tsuji K Sekiya I: Morphological differences during in vitro chondrogenesis of bone marrow- synovium-MSCs and chondrocytes. Lab Invest 2010 90:210-221.
30. Crawford A Frazer A Lippitt JM Buttle DJ Smith T: A case of chondromatosis indicates a synovial stem cell aetiology. Rheumatology (Oxford) 2006 45(12):1529-33.
31. Zhou C Zheng H Seol D Yu Y Martin JA: Gene expression profiles reveal that chondrogenic progenitor cells and synovial cells are closely related. J Orthop Res 2014 32(8):981-8.