Migration, Proliferation, and Differentiation of Cord Blood Mesenchymal Stromal Cells Treated with Histone Deacetylase Inhibitor Valproic Acid
Figure 5
High VPA concentration for longer period downregulates the expression of osteogenic, chondrogenic, and myogenic markers. CB MSC were pretreated with or without (control) 1, 5, or 10 mM VPA for 3 h or 6 h. (a) The VPA was washed off and the cells were incubated in osteogenic and chondrogenic media. Differentiation into osteocytes was assessed after 3 weeks by staining for calcium deposits using Alizarin Red (top panel). Differentiation into chondrocytes was assessed after 3 weeks by staining for proteoglycans using Alcian Blue (bottom panel). (b) The expression of osteogenic markers (osterix and osteocalcin) and chondrogenic marker (collagen type X) was determined by gel-based RT-PCR using GAPDH as internal loading control. (c) The expression of myogenic markers (myogenin and MyoD) was evaluated by qRT-PCR using 18S as internal calibrator. The data is representative of two independent experiments. (d) CB-derived MSC were treated with or without (control) 5 mM VPA for 3 h and induced to differentiate the osteogenic, chondrogenic, and myogenic lineages. The gene expression of collagen type X, osteocalcin, osterix, myogenin, and MyoD was examined by gel-based RT-PCR using GAPDH as internal loading control.
