Characterization of canine bone marrow derived stromal cells

Abstract

In order to facilitate investigations in cellular-based therapies for various neurological disorders, we characterized the phenotypic profile and neural properties of canine bone marrow stromal cells (BMSCs). Bone marrow was aspirated from iliac crests of 5 canine cadavers after euthanasia. BMSCs were isolated by Ficoll density gradient followed by plastic adhesion and expanded. Flow cytometric analysis was performed to evaluate the phenotypic profile of expanded BMSCs. Rapid neural differentiation of BMSCs was induced by dibutyryl cyclic AMP. Neural protein expression was analyzed by western blotting and immunocytochemistry pre and post-neural induction. Flow cytometric analysis revealed a phenotypic profile comparable to human BMSCs (CD34-, CD45-, MHC-II-, CD90+, MHC-I+). Canine BMSCs constitutively expressed β-tubulin ш (early neuronal marker) and astrocyte-specific glial fibrillary acidic protein (GFAP). The level of GFAP expression increased post-induction whereas that of β-tubulin ш remained unchanged. MBP-positive BMSCs were not found under our culture condition. The results suggested that canine BMSCs have neural properties as in other species, and therefore might hold therapeutic benefits for neurological disorders. Studies of BMSC transplantation in spontaneous neurological disorders in dogs may provide important insights into the procedural safety and clinical efficacy as valuable translational animal models. This study was supported by Univeristy of Florida, College of Veterinary Medicine, Consolidated Faculty Research Development Award Grant Competition.