Cultivation and Differentiation of Rat Dental Pulp Stem Cells for Tissue Engineering Purposes

Abstract

Although existence of dental stem cells has been known since 2000, and these are already well characterized, hardly
any data are available regarding rat dental pulp cells. The aim of our study was to isolate, cultivate and differentiate primary
cell cultures, showing stem cell properties, from rat incisor pulp. Cell viability was determined using the WST-1
test. Osteogenic differentiation potential was examined by von Kossa and Alizarin Red staining, and the neurogenic potential
by morphological studies, immunocytochemistry and quantitative PCR. Moreover, the effect of different surface
coatings on neurogenic differentiation was also investigated. We have succeeded in isolating stem cell-containing cell
cultures from the pulp tissue of rat lower incisors and optimized the culture conditions. We have provided evidence that
these cell cultures are capable for differentiating towards both osteogenic and neurogenic lineages. We have developed
a novel neurogenic differentiation protocol, which is more effective than the previous one. Lysine-laminin and ornithinlaminin
coated surfaces were found to be better for rat pulp-cell differentiation. Our results confirm that dental-pulp stem
cells are capable of regenerating injured bone and neural tissue. Our model provides the foundation for the clinical use
of dental pulp stem cells for tissue engineering purposes.