CHARACTERIZATION OF MESENCHYMAL STROMAL CELLS DERIVED OF CANINE DENTAL PULP AND CANINE UMBILICAL CORD

Background Mesenchymal stromal cells (MSC) can be defined as adult, multipotent cells with the ability to differentiate into several cell types. The International Society of Cell Therapy has set minimum criteria for defining MSCs. These MSC can be isolated from different sources such as: dental pulp and umbilical cord. The use of MSC for regenerative medicine for pets has been growing, and an increasing number of studies have contributed to the widespread use of cell therapies in clinical veterinary medicine. Aiming standardization of the MSC used in clinical practice this study isolated, characterized and evaluated the potential of proliferation of MSC derived from dental pulp (cDPSC) and umbilical cord tissue (cUCSC) from dogs. Methods The Local Animal Ethics Committee approved this study (01211/2018). Samples of dental pulp (n=3) and umbilical cord (n=3) were collected. The samples were mechanically dissociated, isolated by enzymatic digestion, and cultivated with IMDM+15% fetal bovine serum. For immunophenotypic characterization, antibodies were used to analyze the expression of MSC markers and to analyze cell viability and apoptosis were used 7-AAD and Annexin-V, respectively. The differentiation was made for adipogenic, osteogenic and chondrogenic lineages. For proliferation assay, 10,000 cells/well were plated in a 6-well plate, enzymatic dissociation was performed every five days and cells were counted in a hemocytometer. Results The cDPSC and cUCSC showed adherence to the plastic and fibroblast-like morphology after culture according to guidelines of the International Society of Cell Therapy (ISCT) for MSC. Both sources showed positivity for CD29 e CD44, reduced expression for CD14, CD45, CD34, and HLA-DR and low mortality and apoptosis rates. In addition, they also shown differentiation potential for the three lineages: osteogenic, adipogenic and chondrogenic. In the third passage, cUCSC showed greater proliferation rates (p=0.0414) compared to cDPSC. Conclusion cDPSC and cUCSC were easily obtained cells with non-invasive collection have demonstrated characteristic of MSC. Both sources can be a safe alternative to cell therapy in dogs.