FIG. 5.
Quantitative polymerase chain reaction (qPCR) analysis of adipogenic and osteogenic marker gene expression levels in MSCs retrieved from culture conditions. MSCs were recovered from 2D (by trypsin) and 3D cultures (by nattokinase) after 14 days of preculture and then subjected to either adipogenic or osteogenic induction media for either an additional 7 or 21 days, respectively. Total RNA was extracted from the cells and subjected to qPCR analysis to assess the expression levels of (A) PPARγ, (B) CEBPα, (C) Runx2, and (D) BGLAP. Statistics were performed on ΔΔCT values and are indicated as shown (**p≤0.01, ***p≤0.001). Collectively, these data showed that MSCs precultured in 3D fibrin gels and subjected to nattokinase extraction were capable of subsequently expressing genes consistent with osteogenic and adipogenic differentiation potentials.