Abstract
Background: Cell-based therapy has achieved good functional recovery for tissue repair. Mesenchymal stemcells (MSCs) exhibit multilineage potential, long-term viability, and capacity for self-renewal. Periosteum-derivedmesenchymal stem cells (PD cells) may be an attractive cell source for tissue engineering because of their easyaccessibility and reduced ethical concerns. Objectives: To isolate and investigate the phenotypic and functional characteristics of mesenchymal stem cellsderived from human periosteum. We also examined the differentiation of PD cells with a trilineage differentiationassay to determine whether they were MSCs.Materials and Methods: Periosteum-derived cells were cultured in osteogenic, chondrogenic or adipogenicmedia to evaluate their multilineage differentiation potential. Adherent fibroblast-like cells were analyzed byflow cytometry for MSC cell surface markers. Differentiation of PD cells into osteogenic, chondrogenic, andadipogenic lineages was also evaluated by von Kossa, Alizarin red, Alcian blue, and oil red O stains, respectively. Expression of mesenchymal stem cell markers were assessed using reverse transcriptase-polymerase chainreaction (RT-PCR) analysis. Results: We successfully isolated and expanded MSCs from human periosteum. Flow cytometry revealed thatPD cells were positive for mesenchymal adhesion cell markers (CD29, CD44, CD90, and CD105) and negative forhematopoietic markers (CD34 and CD45). In osteogenic differentiation, calcium accumulation (positive vonKossa and Alizarin red) and RUNX2, alkaline phosphatase, collagen type I, osteopontin genes were detected.In adipogenic differentiation, the cells displayed oil red O positive and expressed lipoprotein lipase andperoxisome proliferator-activated receptor-gamma (PPAR-?) associated with adipogenesis. The cells grownin chondrogenic conditions were positively stained for Alcian blue and expressed SOX-9. Conclusion: PD cells presented osteogenic, chondrogenic, and adipogenic differentiation abilities in vitro andcould provide an alternative cellular source for tissue repair in clinical applications. Keywords: Adipogenic, chondrogenic, differentiation, mesenchymal stem cells, osteogenic, periosteum
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