Tissue-engineered bone regeneration offers new therapeutic options in the treatment of patients with fractures. Due to the changes in the hormone levels, elderly women are most affected by bone fractures and thus constitute one of the future target groups of cell-based bone therapy. For designing cell-based therapy approaches, a better understanding of individual-dependent variations in bone-derived cells of the host is required. In this study, a simple, high-yield procedure is described for the collection of cells from bone tissue of a high number of elderly women. The cultured cells display stem cell characteristics indicated by the presence of aCD13+, CD44+, CD90+, CD147+, CD14–, CD34–, CD45– and CD144– cell populations and by a stable undifferentiatedphenotype as well as by the ability to proliferate extensivelywhile retaining the potential to differentiate along the osteoblastic lineage even after 27 cell doublings. A high variability in the number of cell-forming units (CFUs) within a donor population of 34 samples, in the morphology within 50 donors, in the expression of alkaline phosphatase within 15 samples and in the responsiveness to BMP-2 was evident, but no age-related correlation could be found. In conclusion, the data indicate that individual variations in cell number, cell morphology and in the osteogenic potential of progenitor cells of the patient may be relevant for a successful treatment of bone fractures in the elderly by cell-based therapy approaches.

Keywords:
Donor variability, Human trabecular bone-derived cells, Mesenchymal stem cells, Osteogenesis, Trabecular bone
1.
Bruder, S.P., D.J. Fink, A.I. Caplan (1994) Mesenchymal stem cells in bone development, bone repair, and skeletal regeneration therapy. J Cell Biochem 56: 283–294.
2.
Bruder, S.P., N. Jaiswal, S.E. Haynesworth (1997) Growth kinetics, self-renewal, and the osteogenic potential of purified human mesenchymal stem cells during extensive subcultivation and following cryopreservation. J Cell Biochem 64: 278–294.
3.
Bruder, S.P., N. Jaiswal, N.S. Ricalton, J.D. Mosca, K.H. Kraus, S. Kadiyala (1998) Mesenchymal stem cells in osteobiology and applied bone regeneration. Clin Orthop 247: S256.
4.
Bruder, S.P., A.A. Kurth, M. Shea, W.C. Hayes, N. Jaiswal, S. Kadiyala (1998) Bone regeneration by implantation of purified, culture-expanded human mesenchymal stem cells. J Orthop Res 16: 155–162.
5.
Cancedda, R., M. Mastrogiacomo, G. Bianchi, A. Derubeis, A. Muraglia, R. Quarto (2003) Bone marrow stromal cells and their use in regenerating bone. Novartis Found Symp 249: 133–143; discussion 143–147, 170–174, 239–241.
6.
Caplan, A.I., S.P. Bruder (2001) Mesenchymal stem cells: Building blocks for molecular medicine in the 21st century. Trends Mol Med 7: 259–264.
7.
Colter, D.C., I. Sekiya, D.J. Prockop (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: 7841–7845.
8.
Ducy, P., R. Zhang, V. Geoffroy, A.L. Ridall, G. Karsenty (1997) Osf2/Cbfa1: A transcriptional activator of osteoblast differentiation. Cell. 89: 747–754.
9.
Egrise, D., D. Martin, A. Vienne, P. Neve, A. Schoutens (1992) The number of fibroblastic colonies formed from bone marrow is decreased and the in vitro proliferation rate of trabecular bone cells increased in aged rats. Bone 13: 355–361.
10.
Glowacki, J. (1995) Influence of age on human marrow. Calcif Tissue Int 56(suppl 1): 50–51.
11.
Groeneveld, E.H., E.H. Burger (2000) Bone morphogenetic proteins in human bone regeneration (review). Eur J Endocrinol 142: 9–21.
12.
Haynesworth, S.E., M.A. Baber, A.I. Caplan (1996) Cytokine expression by human marrow-derived mesenchymal progenitor cells in vitro: Effects of dexamethasone and IL-1 alpha. J Cell Physiol 166: 585–592.
13.
Jaiswal, N., S.E. Haynesworth, A.I. Caplan, S.P. Bruder (1997) Osteogenic differentiation of purified, culture-expanded human mesenchymal stem cells in vitro. J Cell Biochem 64: 295–312.
14.
Katzburg, S., M. Lieberherr, A. Ornoy, B.Y. Klein, D. Hendel, D. Somjen (1999) Isolation and hormonal responsiveness of primary cultures of human bone-derived cells: Gender and age differences. Bone 25: 667–673.
15.
Keating, A., L. Berkahn, R. Filshie (1998) A phase I study of the transplantation of genetically marked autologous bone marrow stromal cells. Hum Gene Ther 9: 591–600.
16.
Kraus, K.H., S. Kadiyala, H. Wotton, A. Kurth, M. Shea, M. Hannan, W.C. Hayes, C.A. Kirker-Head, S. Bruder (1999) Critically sized osteo-periosteal femoral defects: A dog model. J Invest Surg 12: 115–124.
17.
Kuznetsov, S.A., P.H. Krebsbach, K. Satomura, J. Kerr, M. Riminucci, D. Benayahu, P.G. Robey (1997) Single-colony derived strains of human marrow stromal fibroblasts form bone after transplantation in vivo. J Bone Miner Res 12: 1335–1347.
18.
Lazarus, H.M., S.E. Haynesworth, S.L. Gerson, N.S. Rosenthal, A.I. Caplan (1995) Ex vivo expansion and subsequent infusion of human bone marrow-derived stromal progenitor cells (mesenchymal progenitor cells): Implications for therapeutic use. Bone Marrow Transplant 16: 557–564.
19.
Maes, C., P. Carmeliet, K. Moermans, I. Stockmans, N. Smets, D. Collen, R. Bouillon, G. Carmeliet (2002) Impaired angiogenesis and endochondral bone formation in mice lacking the vascular endothelial growth factor isoforms VEGF164 and VEGF188. Mech Dev 111: 61–73.
20.
Majors, A.K., C.A. Boehm, H. Nitto, R.J. Midura, G.F. Muschler (1997) Characterization of human bone marrow stromal cells with respect to osteoblastic differentiation. J Orthop Res 15: 546–557.
21.
Mueller, S.M., J. Glowacki (2001) Age-related decline in the osteogenic potential of human bone marrow cells cultured in three-dimensional collagen sponges. J Cell Biochem 82: 583–590.
22.
Phinney, D.G., G. Kopen, W. Righter, S. Webster, N. Tremain, D.J. Prockop (1999) Donor variation in the growth properties and osteogenic potential of human marrow stromal cells. J Cell Biochem 75: 424–436.
23.
Pittenger, M.F., A.M. Mackay, S.C. Beck, R.K. Jaiswal, R. Douglas, J.D. Mosca, M.A. Moorman, D.W. Simonetti, S. Craig, D.R. Marshak (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284: 143–147.
24.
Quarto, R., M. Mastrogiacomo, R. Cancedda, S.M. Kutepov, V. Mukhachev, A. Lavroukov, E. Kon, M. Marcacci (2001) Repair of large bone defects with the use of autologous bone marrow stromal cells. N Engl J Med 344: 385–386.
25.
Reyes, M., T. Lund, T. Lenvik, D. Aguiar, L. Koodie, C.M. Verfaillie (2001) Purification and ex vivo expansion of postnatal human marrow mesodermal progenitor cells. Blood 98: 2615–2625.
26.
Robey, P.G., J.D. Termine (1985) Human bone cells in vitro. Calcif Tissue Int 37: 453–460.
27.
Rodriguez, J.P., S. Garat, H. Gajardo, A.M. Pino, G. Seitz (1999) Abnormal osteogenesis in osteoporotic patients is reflected by altered mesenchymal stem cells dynamics. J Cell Biochem 75: 414–423.
28.
Sottile, V., C. Halleux, F. Bassilana, H. Keller, K. Seuwen (2002) Stem cell characteristics of human trabecular bone-derived cells. Bone 30: 699–704.
29.
Zuk, P.A., M. Zhu, P. Ashjian, D.A. De Ugarte, J.I. Huang, H. Mizuno, Z.C. Alfonso, J.K. Fraser, P. Benhaim, M.H. Hedrick (2002) Human adipose tissue is a source of multipotent stem cells. Mol Biol Cell 13: 4279–4295.
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2004
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