Isolation and culture of stem cells derived from dental pulp of permanent teeth: a successful method / Isolation and culture of stem cells derived from dental pulp of permanent teeth: a successful method

ABSTRACT

Objective: To establish cultures of cells from the pulp of permanent teeth by the explant method assessing parameters usually presented by stem cells, such as the expression of certain markers and the differentiation ability into osteogenic, adipogenic, and chondrogenic lineages. This study also aimed to assess the expression of ALDH1 (aldehyde dehydrogenase 1) enzyme activity on the isolated cells. Materials and method: The pulp tissue, obtained from wisdom teeth, was placed in a 6-well plate containing proper culture medium, and stored at 37 °C and 5% CO2 for cell proliferation and plastic adherence. Cells were tested for the expression of surface markers and for ALDH1 enzyme activity, by flow cytometry. In addition, cells were assessedfor multi-differentiation potential. Results: The isolated cells showed high expression of CD44 (98.8%), CD73 (100%), and CD90 (97.2%), and moderate expression of STRO-1 (18.4%) and ALDH1 (16.2%), by flow cytometry. Similarly, the cells showed differentiation ability into all three lineages of cells tested. Conclusion: Our results suggest that the explant method - or cell proliferation method - is suitable for the isolation and cultureof stem cells from dental pulp of permanent teeth. The isolated cells may be considered stem cells, based on the current criteria for their characterization, such as plastic adherence, expression of certain markers, and the absence of others, as well as multi-differentiation potential, which showed to be promising for the application in tissue regeneration.