Title: 3396 - Conditioned Medium Assessment from Human Dental Stem Cells Cultures
Lourdes Sitton, Pontificia Universidad Javeriana
Angel Pirela, Pontificia Universidad Javeriana
Camilo Duran, Pontificia Universidad Javeriana
Nelly Roa (Presenter)
Pontificia Universidad Javeriana
Lorenza Jaramillo, Pontificia Universidad Javeriana
Objectives: To evaluate the multidifferentiation potential and cytokines from dental pulp (DPSCs) and periodontal ligament (PLDSCs) stem cells, cultured according to standardized method that allows its potential use as sources of stem cells and conditioned medium in tissue regeneration and/or modulation of oral inflammatory processes.
Methods: This study was approved by the Ethics Committee. Periodontal ligament and dental pulp of third molars were obtained and enzymatically disintegrated and cultured in flasks to establish the primary cultures and to do cell expansion to P6. CD14+ populations were depleted by means of a bounded to anti-CD14 magnetic beads in P1. P0-P6 CD45-/CD34-/CD73+/CD90+/CD105+/CD271+ immunophenotype, proliferation, ability to form colonies, potential to differentiate into multiple cell lines by induction with osteogenic, chondrogenic and adipogenic differentiation medium were evaluated. At P3-P5 the cultures were added with serum-free for 48 and 72 hours, and MCP-1, RANTES, VEGF, EGF, IL-6 were quantified by Luminex MagPix. ANOVA, t-Student and Kruskal Wallis tests were used to analyze the results.
Results: DPSCs and PDLSCs in P1 were 45-55% CD14+. At P2-P6 the cultures were free-CD14+, retained morphology and stem cell immunophenotype with high levels of CD73+/CD90+/CD105+/CD271+ and negative for CD45/CD34, showed positivity to dyes Red Alizarin, Alcian Blue and Oil Red, PDLSC showed higher adipogenic differentiation as compared to the PDSC. In the conditioned medium the cytokines concentration showed statistically significant differences in each passage at 48 and 72 hours, but no between the passages at 48 and 72 hours or between the two sources of stem cells.
Conclusions: These results demonstrated that DPSCs and PDLSCs can be isolated and expanded free-CD14+ and that they can remain free of serum up to 72 hours for the collection of their conditioned medium with high concentrations of mediating factors of cellular and angiogenic recruitment such as MCP-1 and VEGF.
This abstract is based on research that was funded entirely or partially by an outside source:
This study was supported by grant (9064) of Centro de Investigaciones Odontológicas (Pontificia Universidad Javeriana).
The submitter must disclose the names of the organizations with which any author have a relationship, the nature of the relationship, and the clinical or research area involved. The following is submitted: NONE