Cell and Tissue Research

, Volume 327, Issue 3, pp 449–462 | Cite as

Comparison of rat mesenchymal stem cells derived from bone marrow, synovium, periosteum, adipose tissue, and muscle

  • Hideya Yoshimura
  • Takeshi Muneta
  • Akimoto Nimura
  • Akiko Yokoyama
  • Hideyuki Koga
  • Ichiro SekiyaEmail author
Regular Article


Mesenchymal stem cells (MSCs) are increasingly being reported as occurring in a variety of tissues. Although MSCs from human bone marrow are relatively easy to harvest, the isolation of rodent MSCs is more difficult, thereby limiting the number of experiments in vivo. To determine a suitable cell source, we isolated rat MSCs from bone marrow, synovium, periosteum, adipose, and muscle and compared their properties for yield, expansion, and multipotentiality. After two passages, the cells in each population were CD11b (−), CD45 (−), and CD90 (+). The colony number per nucleated cells derived from synovium was 100-fold higher than that for cells derived from bone marrow. With regard to expansion potential, synovium-derived cells were the highest in colony-forming efficiency, fold increase, and growth kinetics. An in vitro chondrogenesis assay demonstrated that the pellets derived from synovium were heavier, because of their greater production of cartilage matrix, than those from other tissues, indicating their superiority in chondrogenesis. Synovium-derived cells retained their chondrogenic potential after a few passages. The Oil Red-O positive colony-rate assay demonstrated higher adipogenic potential in synovium- and adipose-derived cells. Alkaline phosphatase activity was greater in periosteum- and muscle-derived cells during calcification. The yield and proliferation potential of rat MSCs from solid tissues was much better than those from bone marrow. In particular, synovium-derived cells had the greatest potential for both proliferation and chondrogenesis, indicating their usefulness for cartilage study in a rat model.


Mesenchymal stem cells Expansion Chondrogenesis Adipogenesis Calcification Rat (Sprague Dawley, male) 



We are grateful to Kenichi Shinomiya, MD, PhD for supporting our studies, to Yusuke Sakaguchi, MD, PhD for help with the characterization of MSCs, to Izumi Nakagawa for excellent technical assistance, to Miyoko Ojima for expert help with the histology, and to Kelly Johanson, PhD for proofreading.


  1. Allard SA, Maini RN, Muirden KD (1988) Cells and matrix expressing cartilage components in fibroblastic tissue in rheumatoid pannus. Scand J Rheumatol Suppl 76:125–129Google Scholar
  2. Arai F, Ohneda O, Miyamoto T, Zhang XQ, Suda T (2002) Mesenchymal stem cells in perichondrium express activated leukocyte cell adhesion molecule and participate in bone marrow formation. J Exp Med 195:1549–1563PubMedCrossRefGoogle Scholar
  3. Asakura A, Komaki M, Rudnicki M (2001) Muscle satellite cells are multipotential stem cells that exhibit myogenic, osteogenic, and adipogenic differentiation. Differentiation 68:245–253PubMedCrossRefGoogle Scholar
  4. Aubin JE (1999) Osteoprogenitor cell frequency in rat bone marrow stromal populations: role for heterotypic cell-cell interactions in osteoblast differentiation. J Cell Biochem 72:396–410PubMedCrossRefGoogle Scholar
  5. Baksh D, Song L, Tuan RS (2004) Adult mesenchymal stem cells: characterization, differentiation, and application in cell and gene therapy. J Cell Mol Med 8:301–316PubMedGoogle Scholar
  6. Bianco P, Riminucci M, Gronthos S, Robey PG (2001) Bone marrow stromal stem cells: nature, biology, and potential applications. Stem Cells 19:180–192PubMedCrossRefGoogle Scholar
  7. Colter DC, Class R, DiGirolamo CM, Prockop DJ (2000) Rapid expansion of recycling stem cells in cultures of plastic-adherent cells from human bone marrow. Proc Natl Acad Sci USA 97:3213–3218PubMedCrossRefGoogle Scholar
  8. Colter DC, Sekiya I, Prockop DJ (2001) Identification of a subpopulation of rapidly self-renewing and multipotential adult stem cells in colonies of human marrow stromal cells. Proc Natl Acad Sci USA 98:7841–7845PubMedCrossRefGoogle Scholar
  9. De Bari C, Dell’Accio F, Tylzanowski P, Luyten FP (2001a) Multipotent mesenchymal stem cells from adult human synovial membrane. Arthritis Rheum 44:1928–1942PubMedCrossRefGoogle Scholar
  10. De Bari C, Dell’Accio F, Luyten FP (2001b) Human periosteum-derived cells maintain phenotypic stability and chondrogenic potential throughout expansion regardless of donor age. Arthritis Rheum 44:85–95PubMedCrossRefGoogle Scholar
  11. Dezawa M, Takahashi I, Esaki M, Takano M, Sawada H (2001) Sciatic nerve regeneration in rats induced by transplantation of in vitro differentiated bone-marrow stromal cells. Eur J Neurosci 14:1771–1776PubMedCrossRefGoogle Scholar
  12. Dezawa M, Ishikawa H, Itokazu Y, Yoshihara T, Hoshino M, Takeda S, Ide C, Nabeshima Y (2005) Bone marrow stromal cells generate muscle cells and repair muscle degeneration. Science 309:314–317PubMedCrossRefGoogle Scholar
  13. Dobson KR, Reading L, Haberey M, Marine X, Scutt A (1999) Centrifugal isolation of bone marrow from bone: an improved method for the recovery and quantitation of bone marrow osteoprogenitor cells from rat tibiae and femurae. Calcif Tissue Int 65:411–413PubMedCrossRefGoogle Scholar
  14. Friedenstein AJ (1976) Precursor cells of mechanocytes. Int Rev Cytol 47:327–359PubMedGoogle Scholar
  15. Friedenstein AJ, Gorskaja JF, Kulagina NN (1976) Fibroblast precursors in normal and irradiated mouse hematopoietic organs. Exp Hematol 4:267–274PubMedGoogle Scholar
  16. Hofstetter CP, Schwarz EJ, Hess D, Widenfalk J, El Manira A, Prockop DJ, Olson L (2002) Marrow stromal cells form guiding strands in the injured spinal cord and promote recovery. Proc Natl Acad Sci USA 99:2199–2204PubMedCrossRefGoogle Scholar
  17. Ichinose S, Tagami M, Muneta T, Sekiya I (2005) Morphological examination during in vitro cartilage formation by human mesenchymal stem cells. Cell Tissue Res 322:217–226PubMedCrossRefGoogle Scholar
  18. Javazon EH, Colter DC, Schwarz EJ, Prockop DJ (2001) Rat marrow stromal cells are more sensitive to plating density and expand more rapidly from single-cell-derived colonies than human marrow stromal cells. Stem Cells 19:219–225PubMedCrossRefGoogle Scholar
  19. Johnstone B, Hering TM, Caplan AI, Goldberg VM, Yoo JU (1998) In vitro chondrogenesis of bone marrow-derived mesenchymal progenitor cells. Exp Cell Res 238:265–272PubMedCrossRefGoogle Scholar
  20. McKay R (1997) Stem cells in the central nervous system. Science 276:66–71PubMedCrossRefGoogle Scholar
  21. Neubauer M, Fischbach C, Bauer-Kreisel P, Lieb E, Hacker M, Tessmar J, Schulz MB, Goepferich A, Blunk T (2004) Basic fibroblast growth factor enhances PPARgamma ligand-induced adipogenesis of mesenchymal stem cells. FEBS Lett 577:277–283PubMedCrossRefGoogle Scholar
  22. Nishida T, Kubota S, Kojima S, Kuboki T, Nakao K, Kushibiki T, Tabata Y, Takigawa M (2004) Regeneration of defects in articular cartilage in rat knee joints by CCN2 (connective tissue growth factor). J Bone Miner Res 19:1308–1319PubMedCrossRefGoogle Scholar
  23. Peister A, Mellad JA, Larson BL, Hall BM, Gibson LF, Prockop DJ (2004) Adult stem cells from bone marrow (MSCs) isolated from different strains of inbred mice vary in surface epitopes, rates of proliferation, and differentiation potential. Blood 103:1662–1668PubMedCrossRefGoogle Scholar
  24. Phinney DG, Kopen G, Isaacson RL, Prockop DJ (1999) Plastic adherent stromal cells from the bone marrow of commonly used strains of inbred mice: variations in yield, growth, and differentiation. J Cell Biochem 72:570–585PubMedCrossRefGoogle Scholar
  25. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147PubMedCrossRefGoogle Scholar
  26. Prockop DJ (1997) Marrow stromal cells as stem cells for nonhematopoietic tissues. Science 276:71–74PubMedCrossRefGoogle Scholar
  27. Prockop DJ, Gregory CA, Spees JL (2003) One strategy for cell and gene therapy: harnessing the power of adult stem cells to repair tissues. Proc Natl Acad Sci USA 100 (Suppl 1):11917–11923PubMedCrossRefGoogle Scholar
  28. Rodriguez E, Ribot J, Rodriguez AM, Palou A (2004) PPAR-gamma2 expression in response to cafeteria diet: gender- and depot-specific effects. Obes Res 12:1455–1463PubMedCrossRefGoogle Scholar
  29. Sakaguchi Y, Sekiya I, Yagishita K, Ichinose S, Shinomiya K, Muneta T (2004) Suspended cells from trabecular bone by collagenase digestion become virtually identical to mesenchymal stem cells obtained from marrow aspirates. Blood 104:2728–2735PubMedCrossRefGoogle Scholar
  30. Sakaguchi Y, Sekiya I, Yagishita K, Muneta T (2005) Comparison of human stem cells derived from various mesenchymal tissues: superiority of synovium as a cell source. Arthritis Rheum 52:2521–2529PubMedCrossRefGoogle Scholar
  31. Santa Maria L, Rojas CV, Minguell JJ (2004) Signals from damaged but not undamaged skeletal muscle induce myogenic differentiation of rat bone-marrow-derived mesenchymal stem cells. Exp Cell Res 300:418–426PubMedCrossRefGoogle Scholar
  32. Scutt A, Bertram P (1995) Bone marrow cells are targets for the anabolic actions of prostaglandin E2 on bone: induction of a transition from nonadherent to adherent osteoblast precursors. J Bone Miner Res 10:474–487PubMedCrossRefGoogle Scholar
  33. Sekiya I, Colter DC, Prockop DJ (2001) BMP-6 enhances chondrogenesis in a subpopulation of human marrow stromal cells. Biochem Biophys Res Commun 284:411–418PubMedCrossRefGoogle Scholar
  34. Sekiya I, Larson BL, Smith JR, Pochampally R, Cui JG, Prockop DJ (2002a) Expansion of human adult stem cells from bone marrow stroma: conditions that maximize the yields of early progenitors and evaluate their quality. Stem Cells 20:530–541PubMedCrossRefGoogle Scholar
  35. Sekiya I, Vuoristo JT, Larson BL, Prockop DJ (2002b) In vitro cartilage formation by human adult stem cells from bone marrow stroma defines the sequence of cellular and molecular events during chondrogenesis. Proc Natl Acad Sci USA 99:4397–4402PubMedCrossRefGoogle Scholar
  36. Sekiya I, Larson BL, Vuoristo JT, Cui JG, Prockop DJ (2004) Adipogenic differentiation of human adult stem cells from bone marrow stroma (MSCs). J Bone Miner Res 19:256–264PubMedCrossRefGoogle Scholar
  37. Sekiya I, Larson BL, Vuoristo JT, Reger RL, Prockop DJ (2005) Comparison of effect of BMP-2, −4, and −6 on in vitro cartilage formation of human adult stem cells from bone marrow stroma. Cell Tissue Res 320:269–276PubMedCrossRefGoogle Scholar
  38. Shinmei M, Miyauchi S, Machida A, Miyazaki K (1992) Quantitation of chondroitin 4-sulfate and chondroitin 6-sulfate in pathologic joint fluid. Arthritis Rheum 35:1304–1308PubMedGoogle Scholar
  39. Shirasawa S, Sekiya I, Sakaguchi Y, Yagishita K, Ichinose S, Muneta T (2006) In vitro chondrogenesis of human synovium-derived mesenchymal stem cells: optimal condition and comparison with bone marrow-derived cells. J Cell Biochem 97:84–97PubMedCrossRefGoogle Scholar
  40. Simmons DJ, Seitz P, Kidder L, Klein GL, Waeltz M, Gundberg CM, Tabuchi C, Yang C, Zhang RW (1991) Partial characterization of rat marrow stromal cells. Calcif Tissue Int 48:326–334PubMedGoogle Scholar
  41. Valmassy R, Ferguson H (1992) Synovial osteochondromatosis. A brief review. J Am Podiatr Med Assoc 82:427–431PubMedGoogle Scholar
  42. Woodbury D, Schwarz EJ, Prockop DJ, Black IB (2000) Adult rat and human bone marrow stromal cells differentiate into neurons. J Neurosci Res 61:364–370PubMedCrossRefGoogle Scholar
  43. Yokoyama A, Sekiya I, Miyazaki K, Ichinose S, Hata Y, Muneta T (2005) In vitro cartilage formation of composites of synovium-derived mesenchymal stem cells with collagen gel. Cell Tissue Res 322:289–298PubMedCrossRefGoogle Scholar
  44. Yoshida K, Miyauchi S, Kikuchi H, Tawada A, Tokuyasu K (1989) Analysis of unsaturated disaccharides from glycosaminoglycuronan by high-performance liquid chromatography. Anal Biochem 177:327–332PubMedCrossRefGoogle Scholar
  45. Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H, Alfonso ZC, Fraser JK, Benhaim P, Hedrick MH (2002) Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 13:4279–4295PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Hideya Yoshimura
    • 1
  • Takeshi Muneta
    • 1
    • 2
  • Akimoto Nimura
    • 1
  • Akiko Yokoyama
    • 1
  • Hideyuki Koga
    • 1
  • Ichiro Sekiya
    • 3
    Email author
  1. 1.Section of Orthopaedic Surgery, Graduate SchoolTokyo Medical and Dental UniversityTokyoJapan
  2. 2.Center of Excellence Program for Frontier Research on Molecular Destruction and Reconstruction of Tooth and BoneTokyo Medical and Dental UniversityTokyoJapan
  3. 3.Section of Cartilage Regeneration, Graduate SchoolTokyo Medical and Dental UniversityTokyoJapan

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