The inhibitory effects of BMP4 and p21 SiRNA complexes with Exosomes on Wnt signaling Pathway and SH-SY5Y neural stem cell differentiation

Abstract

Our main goal in this study was to investigate the efficiency of exosomes in effective delivering siRNAs targeting P21 and BMP4 in to SH-SY5Y cells. Materials and Methods: Exosomes were isolated from SH-SY5Y cells’ supernatant by using ultracentrifuge based protocols and they were loaded with test siRNAs using ExoFection kit. Afterwards, the effect of siRNAs targeting P21 and BMP-4 on the expression of genes, BMP-4, P21, NeuroD1 and Prox1 in SH-SY5Y cells were assessed by western blot and Real Time PCR. Results: Exosomal markers (CD63 and CD9 ) were confirmed by Flow cytometry . Exosomes’ size was estimated 49.23± 8.43nm by using DLS. Our data showed that exosomes’ cytotoxicity in comparison to Lipofectamine were significantly low. Evaluation of gene expression showed significant increase in Neuro D1 and ProX1 expression level.