A Number of Bone Marrow Mesenchymal Stem Cells but Neither Phenotype Nor Differentiation Capacities Changes with Age of Rats

  • Tokalov, Sergey V. (OncoRay - Center for Radiation Research in Oncology, Medical Faculty Carl Gustav Carus Dresden University of Technology) ;
  • Gruner, Susanne (OncoRay - Center for Radiation Research in Oncology, Medical Faculty Carl Gustav Carus Dresden University of Technology) ;
  • Schindler, Sebastian (OncoRay - Center for Radiation Research in Oncology, Medical Faculty Carl Gustav Carus Dresden University of Technology) ;
  • Iagunov, Alexey S. (Research Laboratory of the Late Radiation Pathology, Central Research Institute of Roentgenology and Radiology) ;
  • Baumann, Michael (OncoRay - Center for Radiation Research in Oncology, Medical Faculty Carl Gustav Carus Dresden University of Technology) ;
  • Abolmaali, Nasreddin D. (OncoRay - Center for Radiation Research in Oncology, Medical Faculty Carl Gustav Carus Dresden University of Technology)
  • Received : 2007.04.11
  • Accepted : 2007.06.19
  • Published : 2007.10.31

Abstract

Bone marrow (BM) derived mesenchymal stem cells (MSC) are pluripotent cells which can differentiate into osteogenic, adipogenic and other lineages. In spite of the broad interest, the information about the changes in BM cell composition, in particularly about the variation of MSC number and their properties in relation to the age of the donor is still controversial. The aim of this study was to investigate the age associated changes in variations of BM cell composition, phenotype and differentiation capacities of MSC using a rat model. Cell populations were characterized by flow cytometry using light scattering parameters, DNA content and a set of monoclonal antibodies. Single cell analysis was performed by conventional fluorescent microscopy. In vitro culture of MSC was established and their phenotype and capability for in vitro differentiation into osteogenic and adipogenic cells was shown. Age related changes in tibiae and femurs, amount of BM tissue, BM cell composition, proportions of separated MSC and yield of MSC in 2 weeks of in vitro culture were found. At the same time, neither change in phenotype no in differentiation capacities of MSC was registered. Age-related changes of the number of MSC should be taken into account whenever MSC are intended to be used for investigations.

Keywords

Acknowledgement

Supported by : BMBF

References

  1. Anisimov, V. N., Khavinson, V. K., Popovich, I. G., Zabezhinski, M. A., Alimova, I. N., et al. (2003) Effect of Epitalon on biomarkers of aging, life span and spontaneous tumor incidence in female Swiss-derived SHR mice. Biogerontology 4, 193−202
  2. Baksh, D., Song, L., and Tuan, R. S. (2004) Adult mesenchymal stem cells: characterization, differentiation, and application in cell and gene therapy. J. Cell Mol. Med. 8, 301−316
  3. Bolliger, A. P. (2004) Cytologic evaluation of bone marrow in rats: indications, methods, and normal morthology. Vet. Clin. Pathol. 33, 58−67
  4. Bos, C., Delmas, Y., Desmouliere, A., Solanilla, A., Hauger, O., et al. (2004) In vivo MR imaging of intravascularly injected magnetically labeled mesenchymal stem cells in rat kidney and liver. Radiology 233, 781−789
  5. Conboy, I. M., Conboy, M. J., Wagers, A. J., Girma, E. R., Weissman, I. L., et al. (2005) Rejuvenation of aged progenitor cells by exposure to a young systemic environment. Nature 443, 760−764
  6. Davis, T. and Kipling, D. (2005) Telomeres and telomerase biology in vertebrates: progress towards a non-human model for replicative senescence and aging. Biogerontology 6, 371− 385
  7. D'Ippolito, G., Schiller, P. C., Ricordi, C., Roos, B. A., and Howard, G. A. (1999) Age-related osteogenic potential of mesenchymal stromal stem cells from human vertebral bone marrow. J. Bone Miner. Res. 14, 1115−1122
  8. Egrise, D., Martin, D., Vienne, A., Neve, P., and Schoutens, K. (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, 335− 361
  9. Endicott, K. M. and Ott, M. (1945) The normal myelogram in albino rats. Anat. Rec. 92, 61−69 https://doi.org/10.1002/ar.1090920209
  10. Jestesen, J., Stenderup, K., Ebbesen, E. N., Mosekilde, L., Steiniche, T., et al. (2001) Adipocyte tissue volume in bone marrow is increased with aging and in patient with osteoporosis. Biogerontology 2, 165−171
  11. Kahn, J. A., Gibbons, R., Perkins, S., and Gazit, D. (1995) Agerelated bone loss: a hypothesis and initial assessment in mice. Clin. Orthop. 313, 69−75
  12. Mets, T., Bekaert, E., and Verdonk, G. (1983) Similarity between in vitro and in vivo cellular aging. Mech. Aging Dev. 22, 71−78
  13. Moerman, E. J., Teng, K., Lipschitz, D. A., and Lecka-Czernik, B. (2004) Aging activates adipogenic and suppress osreogenic programs in mesenchymal marrow stroma/stem cells: the role of $PRAR-{\gamma}2$ transcription factor and $TGF-{\beta}/BMP$ signaling pathways. Aging Cell 3, 379−389
  14. Oswald, J., Boxberger, S., Jorgensen, B., Feldmann, S., Ehninger, G., et al. (2004) Mesenchymal stem cells can be differentiated into endothelial cells in vitro. Stem Cells 22, 377−384
  15. Pittenger, M. F., Mackay, A. M., Beck, S. C., Jaiswal, R. K., Douglas, R., et al. (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284, 143−147 https://doi.org/10.1126/science.284.5411.147
  16. Rickard, D. J., Kassem, M., Hefferan, T. E., Sarkar, G., Spelsberg, T. C., et al. (1996) Isolation and characterization of osteoblast precursor cells from human bone marrow. J. Bone Miner. Res. 11, 312−324
  17. Song, H., Kwon, K., Lim, S., Kang, S. M., Ko, Y. G., et al. (2005) Transfection of mesenchymal stem cells with the FGF-2 gene improves their survival under hypoxic conditions. Mol. Cells 19, 402−407
  18. Stenderup, K., Rosada, C., Jestesen, J., Al-Soubky, T., Dagnaes- Hansen, F., et al. (2004) Aged human bone marrow stromal cells maintaining bone forming capacity in vivo evaluated using an improved method of visualization. Biogerontology 5, 107−118
  19. Stolzing, A. and Scutt, A. (2006) Age-related impairment of mesenchymal progenitor cell function. Aging Cell 5, 213− 214
  20. Tokalov, S. V., Grüner, S., Schindler, S., Wolf, G., Baumann, M., et al. (2007a) Age-related changes in the frequency of mesenchymal stem cells in bone marrow of rats. Stem Cell Dev. 16, (in press)
  21. Tokalov, S. V., Gruner, S., Schindler, S., and Abolmaali, N. D. (2007b) In vitro cultures of rat MSCs in different growth media, Res. J. Biol. Sci. 2, 307−310
  22. Urbich, C., Heeschen, C., Aicher, A., Sasaki, K., Bruhl, T., et al. (2005) Cathepsin L is required for endothelial progenitor cell-induced neovascularization. Nat. Med. 11, 206−213
  23. Vacek, A. (2000) Proliferation activity and number of stromal (CFU-f) and haemopoietic (CFUs) stem cells in bone marrow and spleen of rats of different ages. Acta Vet. 69, 25−31
  24. Xu, C. X., Hendry, J. H., Testa, N. G., and Allen, T. D. (1983) Stromal colonies from mouse marrow: characterization of cell types, optimization of plating efficiency and its effect on radiosensitivity J. Cell Sci. 61, 453−466
  25. Yagunov, A. S., Tokalov, S. V., Chuchlovin, A. B., and Afanassiev, B. V. (1998) Animal studies of residual hematopoietic and immune system injury from low dose/low dose rate radiation and heavy metals, AFRRI, Bethesda