FIG. 4.

Trilineage differentiation of H9-MSCs. H9-MSCs were cultured in the osteogenic differentiation medium for 21 days. The cells were fixed in the plate and stained for mineralized plaques with Alizarin Red. The control culture (A) showed no mineral deposits, whereas the culture in the supplemented differentiation medium showed bright staining of mineralized plaques (B1). (B2) provides a higher magnification view of (B1). Additionally, H9-MSCs were cultured as pellets in the chondrogenic medium for 21 days. The cell pellets were removed, embedded in Optimal Cutting Temperature (OCT) compound, and sectioned. The sections were stained with Alcian Blue for the presence of cartilaginous substrate. The control pellet (C) displayed little dark staining compared to the pellet grown in the chondrogenic medium (D1), which showed bright staining and a more compact morphology. (D2) provides a higher magnification view of (D1) and highlights the more compact association of the cells. H9-MSCs were also cultured in the adipogenic medium for 21 days. The cells were fixed in their plate and stained for the presence of fat vacuoles with oil red O. The control culture showed no positive staining for fat vacuoles (E). The culture maintained in the adipogenic differentiation medium displayed staining for the presence of lipids (F1). (F2) provides a higher magnification view of (F1).