Tomic, Paola (2018) Isolation and morphological characterization of IFP-derived stem-like cells: investigation on their potential role in osteoarthritis. [Laurea specialistica a ciclo unico]
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The extrasynovial adipose tissue, Infrapatellar Fat Pad (IFP), is an emerging player in knee osteoarthritis (OA). Little awareness on origin/function of the stem cells component of the IFP exists. This study aims to isolate/characterize IFP-derived stem cells (IFP-dSCs) from OA patients investigating their role in disease development. Isolated IFP-dSCs morphology and ultrastructure were observed by optical and Transmission Electron Microscope (TEM); expansion potential of cells was assessed by long term culturing. IFP-dSCs vitality and metabolic activity was analyzed. Flow cytometry analysis was performed; in particular, the expression profile was CD73high/FGFR2high/IL-6Rhigh/HLA-ABChigh/HLA-DRhigh/CD90+/VEGFR2+/CD34low/CD44low/IL-1Rlow/CD45-/CD117-. At last IFP-dSCs plasticity was also assessed, evaluating their commitment towards the adipogenic, chondrogenic and endothelial lineages. IFP-dSCs isolation required 14 h; cells were fibroblast-like and typically spindle shaped at low density, showing a greater polygonal nucleus at high density. TEM revealed vesicles in the cytoplasm ascribed to autophagosomes. The replication time was 42.7 (plus;minus) 3.8 h. IFP-dSCs immunophenotype suggested a role in modulation of inflammation. Preliminary data about plasticity revealed the ability in differentiating towards adipogenic and endothelial lineages. Experimental evidence on IFP-dSCs seem to correlate histopathological features of IFP in OA (i.e. thickening of interlobular septa, increase in vascularization and innervation) with the stem cell component.
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