Isolation and characterization of highly osteogenic/cementogenic periodontal ligament mesenchymal stem cell clones

Abstract

Cells derived from the periodontal ligament (PDL) have previously been reported to have stem cell-like characteristics and to play an important part to promote the regeneration of periodontal tissue destroyed by periodontal disease. However, these populations have been heterogeneous, and thus far no specific cellular phenotype has yet been established as being more capable in re-building damage tissues. The aim of presente study will be to isolate highly osteogenic/cementogenic PDL stem cell clones and to delineate their phenotypic and functional characteristics. Using "ring-cloning" technique, cell clones (homogenous populations) will be obtained from three PDL mesenchymal stem cell populations and assessed to: a) osteogenic/cementogenic induction evaluated by von Kossa/alizarin red staining and expression of RUNX2, ALP and OCN, b) adipogenic induction assessed by oil red O staining and PPARgama2 and LPL gene expression, c) proliferation rate and viability by MTS, d) immunostaining for STRO-1, e) screening of surface markers assessed by gene expression (RNAseq technique), and mass spectometry of protein . After identification of surface markers expressed by osteogenic/cementogenic clone cells, specific assays will be perfomed to assess the signaling pathway by these markers promote cell differentiation. (AU)