Research Article
A Small-Sized Population of Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells Shows High Stemness Properties and Therapeutic Benefit
Figure 2
Stem cell characterization of differentially-sized populations. (a) Cell morphology was analyzed by microscopy ( μm). Three populations were observed with spindle-shape morphology. (b) During incubation in specialized induction media, multilineage differentiation was evaluated by staining for typical lineage markers. Among the three populations, osteogenic cells were analyzed based on bone-type ALP (upper panel). The picture with full range is shown in the boxed insert. Chondrogenic cells accumulated sulfated proteoglycan that stained positively for safranin O (middle panel). Adipogenic cells showed enhanced lipid vacuoles within the cytoplasm, which were stained with oil red O (lower panel; μm). Nuclei were counterstained with hematoxylin. (c) Stemness genes (Oct4, Nanog) were quantified by qPCR. The expression levels of all genes were normalized to those of β-actin, and expression levels were normalized to those observed in heterogeneous cells, defined as 1-fold expression (, ; , ). (d) The confluence of the three cell populations was determined based on adherence to the culture flask at different culture times (, ; , ).
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