Effects of Folic acid on stem cell proliferation and differentiation derived from posterior neural tube of Pax3 mutant (Splotch) mouse

Abstract

Periconceptional maternal folic acid (FA) supplementation has been shown to be extremely effective in reducing the occurrence of neural tube defects (NTDs). However, little is known about the anti‐teratogenic mechanism of FA in neural tube anomalies. The proliferation or differentiation of neural stem cells (NSCs) can be influenced by various maternal factors, leading to alteration of the cell‐fate of the neural crest. Here we try to elucidate how folic acid can influence on stem cell proliferation and differentiation in Pax3 mutant (Splotch) mouse embryo. By using neurosphere culture derived from posterior neural tube of E10.5 mouse embryos, wild type (WT) and Splotch homozygote (Sp−/−), we observed colony forming ability that could be extremely increased by folic acid treatment on Sp−/− that caused neural tube defect in 100% whereas only few colonies could be seen without treatment. Those rescued sphere colonies showed definite immunoreactivity to stem cell, e.g. Oct3/4, Sox2, CD133, nestin and alkaline phosphatase suggesting they were pluripotent progenitor‐like cell. After plating those spheres on Matrigel, cells that migrated out from spheres and differentiated could express GFAP, Brn3a, β‐‡V‐tubulin, O4 and synaptogenesis markers, e.g. Snap25 or synaptotagmin. Thus this study suggested that folic acid could play an important role on stem cell proliferation and differentiation.