Gingival fibroblasts (GFs) that exhibit characteristics similar to adult stem cells are also known as gingival mesenchymal stem cells (GMSCs). These cells can be isolated using either tissue explants or enzymatic digestion, however it is unknown whether the isolation method influences the GMSCs potential for differentiation. In addition, no specific mesenchymal stem cell (MSC) marker has been reported to identify and distinguish GMSCs from GFs. Recently, the cell surface molecule known as

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... r of differentiation (CD) 146 has been identified as a potential MSC cell surface marker. Objective: To investigate the differentiation potential of GMSCs based on CD146 expression. Hypothesis: CD146 is the MSC surface marker that identifies GMSCs among a population of GFs regardless of the isolation method employed. Materials and methods: GFs were isolated by two techniques: tissue explants or enzymatic digestion. GFs were cultured and expanded in a medium containing fibroblast growth factor 2 (FGF-2), and magnetically sorted according to CD146 expression. Four experimental groups were collected: CD146 low and CD146 high from explant-derived isolation and CD146 low and CD146 high from digestion-derived isolation. Each group of cells was expanded and then tested for stem cell markers using flow cytometry before it was subjected to osteogenic and chondrogenic differentiation. Multilineage differentiation outcome was tested after 21 days using histology, immunofluorescence, real-time quantitative PCR (qPCR), and glycosaminoglycan (GAG) to DNA ratio (GAG/DNA) assays.