Abstract
Bone marrow stromal cells (BMSCs) have gained considerable attention as a potential source for cell transplantation therapies for a variety of diseases due to their accessibility, proliferative capacity, and multilineage differentiation properties. Canine BMSCs have been shown to contribute to regeneration of osseous tissues, but knowledge about their biology is currently limited. In the present study, we investigated the frequency of adult canine BMSCs in bone marrow, morphological features, growth kinetics, and osteogenic as well as adipogenic differentiation properties in vitro. Our data suggest that adult canine bone marrow contains approximately one BMSC in every 2.38 × 104 bone marrow mononucleated cells (0.0042 ± 0.0019%, n = 5). Primary BMSC cultures consisted of morphologically heterogeneous adherent cell populations from which spindle-shaped cells grew and became the predominant cell type. Growth kinetics patterns were dependent on the initial cell seeding densities, resulting in the highest fold increase at lower cell density. In the presence of osteogenic and adipogenic inducers, primary BMSCs underwent morphological and phenotypic changes characteristic of osteogenic and adipogenic differentiation, respectively. This study provides insights into basic characterization of adult canine BMSCs.
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Anselme K., et al. In vitro control of human bone marrow stromal cells for bone tissue engineering. Tissue Eng 8: 941–953; 2002. doi:10.1089/107632702320934047
Arinzeh T. L., et al. Allogeneic mesenchymal stem cells regenerate bone in a critical-sized canine segmental defect. J Bone Joint Surg Am 85-A: 1927–1935; 2003.
Arnhold S. J., et al. Isolation and characterization of bone marrow-derived equine mesenchymal stem cells. Am J Vet Res 68: 1095–1105; 2007. doi:10.2460/ajvr.68.10.1095
Bruder S. P., et al. 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; 1997.
Bruder S. P., et al. The effect of implants loaded with autologous mesenchymal stem cells on the healing of canine segmental bone defects. J Bone Joint Surg Am 80: 985–996; 1998.
Caplan A. I., et al. The development of embryonic bone and cartilage in tissue culture. Clin Orthop Relat Res. 102: 243–263; 1983.
Chen L. B., et al. Differentiation of rat marrow mesenchymal stem cells into pancreatic islet beta-cells. World J Gastroenterol 10: 3016–3020; 2004.
Chen Y., et al. In vitro differentiation of mouse bone marrow stromal stem cells into hepatocytes induced by conditioned culture medium of hepatocytes. J Cell Biochem 102: 52–63; 2007. doi:10.1002/jcb.21275
Colter D. C., et al. Rapid expansion of recycling stem cells in cultures of plastic-adherent cells from human bone marrow. Proc Natl Acad Sci U S A 97: 3213–3218; 2000. doi:10.1073/pnas.070034097
Deng J., et al. Mesenchymal stem cells spontaneously express neural proteins in culture and are neurogenic after transplantation. Stem Cells 24: 1054–1064; 2006. doi:10.1634/stemcells.2005-0370
D’Ippolito G., et al. Marrow-isolated adult multilineage inducible (MIAMI) cells, a unique population of postnatal young and old human cells with extensive expansion and differentiation potential. J Cell Sci 117: 2971–2981; 2004. doi:10.1242/jcs.01103
Friedenstein A. J., et al. Fibroblast precursors in normal and irradiated mouse hematopoietic organs. Exp Hematol 4: 267–274; 1976.
Gindraux F., et al. Human and rodent bone marrow mesenchymal stem cells that express primitive stem cell markers can be directly enriched by using the CD49a molecule. Cell Tissue Res 327: 471–483; 2007. doi:10.1007/s00441-006-0292-3
Haig D. M., et al. The in-vitro detection and quantitation of ovine bone marrow precursors of multipotential colony-forming cells. J Comp Pathol 111: 73–85; 1994. doi:10.1016/S0021-9975(05)80113-6
Izadpanah R., et al. Characterization of multipotent mesenchymal stem cells from the bone marrow of rhesus macaques. Stem Cells Dev 14: 440–451; 2005. doi:10.1089/scd.2005.14.440
Kadiyala S., et al. Culture expanded canine mesenchymal stem cells possess osteochondrogenic potential in vivo and in vitro. Cell Transplant 6: 125–134; 1997. doi:10.1016/S0963-6897(96)00279-5
Kamishina H., et al. Expression of neural markers on bone marrow-derived canine mesenchymal stem cells. Am J Vet Res 67: 1921–1928; 2006. doi:10.2460/ajvr.67.11.1921
Kopen G. C., et al. Marrow stromal cells migrate throughout forebrain and cerebellum, and they differentiate into astrocytes after injection into neonatal mouse brains. Proc Natl Acad Sci U S A 96: 10711–10716; 1999. doi:10.1073/pnas.96.19.10711
Lee K. D., et al. In vitro hepatic differentiation of human mesenchymal stem cells. Hepatology 40: 1275–1284; 2004. doi:10.1002/hep.20469
Lennon D. P., et al. A chemically defined medium supports in vitro proliferation and maintains the osteochondral potential of rat marrow-derived mesenchymal stem cells. Exp Cell Res 219: 211–222; 1995. doi:10.1006/excr.1995.1221
Li T. S., et al. The safety and feasibility of the local implantation of autologous bone marrow cells for ischemic heart disease. J Card Surg 18Suppl 2: S69–75; 2003. doi:10.1046/j.1540-8191.18.s2.3.x
Martin D. R., et al. Isolation and characterization of multipotential mesenchymal stem cells from feline bone marrow. Exp Hematol 30: 879–886; 2002. doi:10.1016/S0301-472X(02)00864-0
Meirelles Lda S.; Nardi N. B. Murine marrow-derived mesenchymal stem cell: isolation, in vitro expansion, and characterization. Br J Haematol 123: 702–711; 2003. doi:10.1046/j.1365-2141.2003.04669.x
Memon I. A., et al. Combined autologous cellular cardiomyoplasty with skeletal myoblasts and bone marrow cells in canine hearts for ischemic cardiomyopathy. J Thorac Cardiovasc Surg 130: 646–653; 2005. doi:10.1016/j.jtcvs.2005.02.024
Nadri S.; Soleimani M. Isolation murine mesenchymal stem cells by positive selection. In Vitro Cell Dev Biol Anim 43: 276–282; 2007. doi:10.1007/s11626-007-9041-5
Phinney D. G., et al. 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: 570–585; 1999.
Pittenger M. F., et al. Multilineage potential of adult human mesenchymal stem cells. Science 284: 143–147; 1999. doi:10.1126/science.284.5411.143
Sekiya I., et al. 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: 530–541; 2002. doi:10.1634/stemcells.20-6-530
Shi Q., et al. Evidence for circulating bone marrow-derived endothelial cells. Blood 92: 362–367; 1998.
Silva G. V., et al. Mesenchymal stem cells differentiate into an endothelial phenotype, enhance vascular density, and improve heart function in a canine chronic ischemia model. Circulation 111: 150–156; 2005. doi:10.1161/01.CIR.0000151812.86142.45
Stamm C., et al. Autologous bone-marrow stem-cell transplantation for myocardial regeneration. Lancet 361: 45–46; 2003. doi:10.1016/S0140-6736(03)12110-1
Sun S., et al. Isolation of mouse marrow mesenchymal progenitors by a novel and reliable method. Stem Cells 21: 527–535; 2003. doi:10.1634/stemcells.21-5-527
Tropel P., et al. Isolation and characterisation of mesenchymal stem cells from adult mouse bone marrow. Exp Cell Res 295: 395–406; 2004. doi:10.1016/j.yexcr.2003.12.030
Vidal M. A., et al. Cell growth characteristics and differentiation frequency of adherent equine bone marrow-derived mesenchymal stromal cells: adipogenic and osteogenic capacity. Vet Surg 35: 601–610; 2006. doi:10.1111/j.1532-950X.2006.00197.x
Volk S. W., et al. Effects of osteogenic inducers on cultures of canine mesenchymal stem cells. Am J Vet Res 66: 1729–1737; 2005. doi:10.2460/ajvr.2005.66.1729
Vulliet P. R., et al. Intra-coronary arterial injection of mesenchymal stromal cells and microinfarction in dogs. Lancet 363: 783–784; 2004. doi:10.1016/S0140-6736(04)15695-X
Xu C. X., et al., Stromal colonies from mouse marrow: characterization of cell types, optimization of plating efficiency and its effect on radiosensitivity. J Cell Sci 61: 453–466; 1983.
Yoshimura H., et al. Comparison of rat mesenchymal stem cells derived from bone marrow, synovium, periosteum, adipose tissue, and muscle. Cell Tissue Res 327: 449–462; 2007. doi:10.1007/s00441-006-0308-z
Zangi L., et al. High-yield isolation, expansion, and differentiation of rat bone marrow-derived mesenchymal stem cells with fibrin microbeads. Tissue Eng 12: 2343–2354; 2006. doi:10.1089/ten.2006.12.2343
Zhang Y., et al. In vitro chondrogenic phenotype differentiation of bone marrow-derived mesenchymal stem cells. Journal of Huazhong University Science of Technology and Medical Science 24: 275–278; 2004.
Zohar R., et al. Characterization of stromal progenitor cells enriched by flow cytometry. Blood 90: 3471–3481; 1997.
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This study was supported by Shands HealthCare.
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Kamishina, H., Farese, J.P., Storm, J.A. et al. The frequency, growth kinetics, and osteogenic/adipogenic differentiation properties of canine bone marrow stromal cells. In Vitro Cell.Dev.Biol.-Animal 44, 472–479 (2008). https://doi.org/10.1007/s11626-008-9137-6
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DOI: https://doi.org/10.1007/s11626-008-9137-6