Enhanced neuronic differentiation of neural stem cells by SHH gene-engineered EMSCs co-culture in vitro

Abstract

Spinal cord injury is a severe central nervous system disease. Neural crest-derived ectoderm mesenchymal stem cells (EMSCs) have become an important tool in treating SCI owing to their excellent neuronal differentiation efficiency and promising paracrine effects. Sonic hedgehog (shh) was proven to promote both the proliferation and chemotaxis of NSCs and induce their differentiation into mature neurons and the formation of synapses. In this study, we examined the neuronal differentiation induced by SHH-engineered EMSCs in a co-culture system. The levels of the neuronal markers, tubulin beta 3 class III and growth-associated protein 43, were higher in NSCs in the SHH-EMSC co-culture group, and those of the astrocytic marker glial fibrillary acidic protein (GFAP) were lower, compared with the mono and EMSCs-NSCs groups The expression of GAP43, GFAP, MBP, and NF200 was also confirmed by western blot analysis. Most importantly, the upregulation of con43, con47, smoothen, and integrin might reveal the construction of the gap junction during the overexpression of SHH in EMSCs. Overall, our findings indicate that SHH-EMSCs might facilitate the neuronal differentiation of NSCs via the gap junction formation.