Title: 2836 - Comparative Characterization of Stem Cells from Lewis Rats
Lorenza Jaramillo (Presenter)
Pontificia Universidad Javeriana
Angel Pirela, Pontificia Universidad Javeriana
Nelly Roa, Pontificia Universidad Javeriana
Camilo Duran, Pontificia Universidad Javeriana
Objectives: Compare the in vitro characteristics of stem cells from bone marrow (BMSC) and dental pulp (PDSC) from Lewis rats, to be used in further tissue engineering studies and regenerative medicine.
Methods: With the approval of the Institutional Animal Care and Use Committee of Javeriana University, 10 Lewis rats were euthanized and its posterior extremities and incisor teeth were dissected for P0-P6 from BMSC and DPSC cultures. CD45-, CD90+, CD71+, CD106+, and CD146+ immunophenotype were determined by flow cytometry, and the potential for differentiation into the osteogenic, chondrogenic and adipogenic lineage was tested with culture medium supplemented with induction molecules to these lineages. At P4-P5 the cultures were added with serum-free α-MEM for 48 and 72 hours, to collect the medium conditioned and their concentration of MCP-1, RANTES, VEGF, EGF, IL-6 were determinate through the commercial panel (MerckMillipore RECYTMAG-65K). ANOVA, t-Student and Kruskal Wallis tests were used to analyze the results.
Results: BMSC and DPSC after several culture passages retain their morphology, typical characteristics, and stem cell immunophenotype. Both types of cells have the potential for osteogenic, chondrogenic and adipogenic differentiation, which increases in passages with higher cellular purity and in which the rate of proliferation is also increased. In BMSC a highly positive expression was found for the CD90 and CD29 markers, whereas for the PDSC the expression of CD146 and CD90 was positive without differences with the BMSC. PDSC are 1.5 times more capable of establishing colony forming units and twice the rate of proliferation than BMSC. In the conditioned medium, no IL-6 and EGF were detected, the concentrations of MCP-1 and RANTES were very low and only VEGF showed to be significantly higher in P5 and 72h in DPSC.
Conclusions: PDSC exhibited higher proliferative and phosphatase alkaline activity and in their media conditioned showed a higher level of VEGF concentration in compared with BMSC, thus PDSC may be a useful cell source for application in regeneration studies.
This abstract is based on research that was funded entirely or partially by an outside source:
This study was supported by grant (7358) of Pontificia Universidad Javeriana Vice-Rectory.
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