The Thai Journal of Veterinary Medicine


Mesenchymal stem cells (MSCs) are multipotent cells that have characteristics of self-renewal anddifferentiation into various specific cell types, in particular mesodermal lineages. This study aimed at isolating,identifying and examining the differentiation capability of canine MSCs. Bone marrow aspirates were obtained from4 dogs. Putative MSCs were then cultured in MSC medium and subpassaged. At the 3rd to 5th passages, MSCs wereexamined for their morphology and doubling time. Two cell lines were examined for the expression of CD 34, CD 44and CD 90, using flow cytometry. The in vitro differentiation of these MSCs into mesodermal lineages (bone, cartilageand adipose tissues) and ectodermal lineage (neuron) was performed using osteogenic, chondrogenic, adipogenic andneurogenic media, respectively. Histological examinations (Von Kossa and alcian blue staining) and mRNAexpressions (GLA and COL1A1) were used to examine the bone and cartilage differentiation, while Oil red O stainingwas used to determine adipogenic differentiation. Plastic-adhered MSCs had high potential for cell division, with a mean doubling time of 35.4±9.3 hours.These fibroblast-like MSCs expressed MSCs markers (CD 44 and CD 90), while fewer than 5% of these MSCs weretested positive to a hemopoietic stem cell marker (CD34). Based on the histological examinations and geneexpressions, these cells demonstrated the ability to differentiate into bone, cartilage and adipose tissues. Inconclusion, MSCs can be isolated from canine bone marrow and these cells are capable of in vitro differentiation intospecific mesodermal lineages.



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