Fig. 6. Effect of Has2 deficiency on cartilage neogenesis in alginate bead cultures.
Matrix assembly and retention in 3-D cultures were examined using RCS-Cas9 WT cells and RCS-Cas9 Has2 KO clones 3 and 7 as labeled. Upper six Panels: Cells were cultured in alginate beads for 7 days, and paraffin sections of the beads were stained with Toluidine Blue. Lower magnification images (10× objective lens) are shown in the upper row; higher magnification views (40× objective) in second row. Sections were stained with biotin-HABP revealing HA in the matrix as a brown stain in rows three, four and five, shown at low (4× objective), medium (20× objective) and higher (40× objective) magnification. Safranin O/fast green staining of alginate bead sections are shown in the bottom row (40× objective).