Isolation, expansion and Characterization of Canine Adipose derived Mesenchymal Stem cells (cADMSCs) from Omental tissue

Mesenchymal stem cells from omental fat were harvested from twelve adult clinically healthy dogs for isolation, expansion and characterization of canine Adipose derived Mesenchymal Stem cells (cADMSCs) in-vitro. Collected tissue samples were weighed and digested using collagenase enzyme to isolate cADMSCs. Cell yield and viability of the cells were calculated by using trypan blue exclusion test. Viable cells were seeded at a density of one million cells per T-25 culture flask and incubated in DMEM-HG, containing 10 per cent FBS, at 37°C under 5 per cent CO? tension. In the present study, the amount of cADMSCs collected in SVF was 3.46 X 106 cells per gram of omental adipose tissue. Flow cytometry analysis revealed that only 5 per cent cells were positive for CD34. Cells harvested at passage 3 were maintained in differentiation medium for an optimal time period of 21 days and their differentiation into chondrocytes and was observed by histochemical staining. Differentiated cells were positive for Toluidine blue with distinct cell morphology. These findings suggested that cADMSCs as a reliable source of adult stem cells which can differentiate into mesodermal lineages.