Figure 4

Human cadaver mesenchymal stromal/stem cell mesengenic potential. (A) Control human cadaver mesenchymal stromal/stem cells (hC-MSCs) did not display cytoplasm lipid drops. (B) Oil Red O stained adipocytic multivacuolar cells in red. (A), (B) Scale bars = 10 μm. (C) Transmission electron microscopy (TEM) showed multiple lipid vacuoles and small dense mitochondria in the cytoplasm. L, lipid droplets; M, mitochondria. Scale bar = 2 μm. (D) Reverse transcriptase polymerase chain reaction of peroxisome proliferator-activated receptor gamma (PPARγ) expression. β-Microglobulin was used as the housekeeping gene. (E) Control hC-MSCs did not display calcium deposition in the extracellular matrix. (F) Alizarin Red stained calcium deposits. (E), (F) Scale bars = 10 μm. (G) TEM confirmed the presence of osteoid matrix and needle-shaped hydroxyapatite crystals (arrow). Scale bar = 2 μm. (H) Gene expression analysis of Osteocalcin, Osteopontin and RUNX-2. β-Microglobulin was used as the housekeeping gene. (I) Control hC-MSCs did not display proteoglycan-rich extracellular matrix. (J) Alcian Blue stained proteoglycan-rich extracellular matrix. (K) Glycogen inclusions (arrow) stained by PAS staining with and without diastase pretreatment. (I), (J), (K) Scale bars = 10 μm. (L) Human collagen type II immunostaining positive in the extracellular matrix. Scale bar = 100 μm. (M) TEM analysis revealed proteoglycans adherent to the cell membrane (arrows). Scale bar = 2 μm. (N) Molecular analysis of type II collagen transcript expression. β-Microglobulin was used as the housekeeping gene. (O) Control hC-MSCs did not display contractile filaments. (P) TEM analysis revealed peripherally arranged contractile filaments, dense bodies, glycogen deposits (*) and profiles of rough endoplasmic reticulum. (Q) Elastic lamellae in the extracellular matrix (arrow). O), (P), (Q) Scale bars = 2 μm. Matrigel assay in the absence (R) and presence (S) of vascular endothelial growth factor (VEGF; 50 ng/ml for 7 days) after 6 hours. (R), (S) Scale bars = 10 μm. (T), (U) Flow cytometry analysis for von Willebrand factor (vWF) and CD31 expression in hC-MSCs cultured in the absence and in the presence of VEGF. Uninduced cells are presented as filled black histograms, differentiated cells as white histograms.