Isolation, characterization and transduction of canine bonemarrow-derived mesenchymal stem cells (cBM-MSCs)

BACKGROUND: Stem cell therapy in small animal medicine is still in its infancy and few in vitro and in vivo research projects regarding animal Mesenchymal Stem Cells (MSCs) have been carried out. On the other hand, Cell tracking is the first step of the cell-based therapies and is essential to recognize cell fate posttransplantation. OBJECTIVES: The aim of this study was to isolate, characterize, and transduce cBM-MSCs. METHODS: Canine Bone Marrow-derived Mesenchymal Stem Cells (cBMMSCs) were isolated from bone marrow of dogs and characterized based on morphology, differentiation capacities,and surface marker expressions. For the first time, we labeled cBM-MSCs by GFP-encoding lentiviral vector to track them. RESULTS: cBM-MSCs were successfully isolated and proliferated. Morphologically, these cells were similar to other MSCs fro m other sources and species and were able todifferentiate into osteocytes and adipocytes. cBM-MSCs expressed surface marker CD44 but were not able to express CD34. Approximately, 70% of cells efficaciously expressed GFPafter labeling; CONCLUSIONS:We found that GFP labeling is an easy and effective technique to track transplanted cBMMSCs.Our results also provide fundamental information about canine BM-MSCs in order to use in veterinary medicine.

Keywords:

canine Bone Marrow-derivedMesenchymal Stem Cells (cBMMSCs) , characterization , greenfluorescent protein (GFP) , transduction