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

Figure 5

Early radial glial differentiation of MMC-AFSCs and healthy AFSCs. (a, b) IF analysis of MMC-AFSCs and healthy AFSCs and at day 15 of neural induction, exposed or not with MTX and VPA, showing positive cells for the early radial glial marker BLBP (green) and the neural crest stem cell migration marker PAX3 (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 BLBP⁺ 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 Healthy + MTX, Healthy versus Healthy + VPA, Healthy versus MMC, MMC versus MMC + MTX, and MMC versus MMC + VPA. (d) Enumeration of PAX3⁺ cells in neural-derived healthy AFSCs and MMC-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 MMC, MMC versus MMC + VPA, and MMC + MTX versus MMC + VPA.