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

Figure 4

Proliferative and immature 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 Ki67 (green) and the immature neural differentiation marker βIII-tubulin (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 βIII-tubulin⁺ cells in neural-derived MMC-AFSCs and healthy AFSCs at day 15 of neural induction, exposed or not with MTX and VA, shown as percentage. and values are indicated as mean ± SD. NS, not significant. Healthy + VPA versus MMC + VPA; Healthy versus MMC, MMC versus MMC + MTX, and MMC versus MMC + VPA. (d) Enumeration of Ki67⁺ 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 versus MMC, Healthy versus Healthy + MTX, Healthy versus Healthy + VPA, MMC versus MMC + MTX, and MMC versus MMC + VPA.