Expression of neural markers on bone marrow-derived canine mesenchymal stem cells.
OBJECTIVE
To evaluate cell surface markers of bone marrow-derived canine mesenchymal stem cells (MSCs) by use of flow cytometric analysis and determine whether canine MSCs express proteins specific to neuronal and glial cells.


METHODS
Bone marrow aspirates collected from iliac crests of 5 cadavers of young adult dogs.


METHODS
Flow cytometric analysis was performed to evaluate cell surface markers and homogeneity of third-passage MSCs. Neural differentiation of canine MSCs was induced by use of dibutyryl cAMP and methyl-isobutylxanthine. Expressions of neuronal (beta III-tubulin) and glial (glial fibrillary acidic protein [GFAP] and myelin basic protein) proteins were evaluated by use of immunocytochemical and western blot analyses before and after neural differentiation.


RESULTS
Third-passage canine MSCs appeared morphologically homogeneous and shared phenotypic characteristics with human and rodent MSCs. Immunocytochemical and western blot analyses revealed that canine MSCs constitutively expressed beta III-tubulin and GFAP. After induction of neural differentiation, increased expression of GFAP was found in all samples, whereas such change was inconsistent in beta III-tubulin expression. Myelin basic protein remained undetectable on canine MSCs for these culture conditions.


CONCLUSIONS
Canine bone marrow-derived mononuclear cells yielded an apparently homogeneous population of MSCs after expansion in culture. Expanded canine MSCs constitutively expressed neuron or astrocyte specific proteins. Furthermore, increases of intracellular cAMP concentrations induced increased expression of GFAP on canine MSCs, which suggests that these cells may have the capacity to respond to external signals. Canine MSCs may hold therapeutic potential for treatment of dogs with neurologic disorders.
Kamishina
Hiroaki
H
Department of Small Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL 32610, USA.
Deng
Jie
J
Oji
Takashi
T
Cheeseman
Jennifer A
JA
Clemmons
Roger M
RM
D000818:Animals; D015153:Blotting, Western; D001854:Bone Marrow Cells / Q000378:metabolism*; D004285:Dogs / Q000378:metabolism*; D005434:Flow Cytometry; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism*; D007150:Immunohistochemistry; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D004676:Myelin Basic Protein / Q000378:metabolism*; D014404:Tubulin / Q000378:metabolism*
D005904:Glial Fibrillary Acidic Protein; D004676:Myelin Basic Protein; D014404:Tubulin