Methotrexate and Valproic Acid Affect Early Neurogenesis of Human Amniotic Fluid Stem Cells from Myelomeningocele

Figure 3

Early neural differentiation potential of MMC-AFSCs and healthy AFSCs. (a, b) IF analysis of MMC-AFSCs and healthy AFSCs at day 15 of neural induction, exposed or not with MTX and VPA, showing positive cells for early neural markers SOX2 (green) and NESTIN (red). Representative images of healthy AFSCs (clone number 1) and MMC-AFSCs (clone number 5) were shown for all healthy and MMC lines. Nuclei were counterstained with Hoechst (blue). , scale bar = 100 μm. (c) Enumeration of SOX2⁺ cells in neural-derived MMC-AFSCs and healthy AFSCs at day 15 of neural induction, exposed or not with MTX and VPA, shown as percentage. and values are indicated as mean ± SD. Healthy versus Healthy + MTX, Healthy versus Healthy + VPA, Healthy + MTX versus Healthy + VPA, Healthy versus MMC, MMC versus MMC + MTX, MMC versus MMC + VPA, Healthy + MTX versus MMC + MTX, and Healthy + VPA versus MMC+VPA. (d) Enumeration of NESTIN⁺ cells in neural-derived MMC-AFSCs and healthy AFSCs at day 15 of neural induction, exposed or not with MTX and VPA, shown as percentage. and values are indicated as mean ± SD. NS, not significant. Healthy + VPA versus MMC + VPA; Healthy versus Healthy + MTX; Healthy + MTX versus MMC+MTX, Healthy + MTX versus Healthy + VPA, and MMC + MTX versus MMC + VPA; and Healthy versus Healthy + VPA, MMC versus MMC + MTX, and MMC versus MMC + VPA.