Regenerative medicine has the potential to replace tissues and organs damaged using stem cells (SCs) and their derivatives. Mesenchymal stromal cells (MSCs) are adult cells, multipotent and functionally defined as having self-renewal capacity, differentiation capacity in several cell lines and extensive paracrine and immunomodulatory activity. It has been described the presence of different types of MSCs populations in teeth. Dental pulp-derived SCs (DPSCs) are an important source of SC for cell therapy, can easily be collected and presents a low risk for the patient besides the great availability of material (e.g.: third molar). DPSCs have differentiation potential in multiple lineages like MSCs obtained from other tissues. The use of MSCs on a large scale in the research depends on the quality of the cells that will promote the regeneration of an injured tissue. An important aspect of standardization is the isolation method. The isolated and cultivated DPSCs present fibroblast-like morphology, plastic adherence and demonstrate the immunophenotypic profile suggested by International Society for Cellular Therapy. DPSCs derive from neural crest and have the potential to differentiate into neurons and promote the survival of neuronal cells from the injured site. The characteristics (differentiation and surface markers) presented by DPSCs are very similar with stem cells of other tissues. However, for DPSC to be used on a large scale in regenerative medicine it is necessary to standardize the isolation methodology.