Extracellular vesicles analysis and relationship in ADSC neuronal differentiation - (a) Microscopy of transmission of small vesicles from day 3 of co-culture (Exo-CC D3) and day 7 of co-culture (Exo-CC D7). (b) Particle diameter (nm) distribution of the isolated particles from control and co-culture (CC) media isolated on day three (D3) and day seven (D7) (Small EVs-Exo) according to nano-tracking analysis. (c) Extracellular vesicles concentration according to nano-tracking analysis. (d) The western blotting analysis of ALIX, HPS70, CD63, CD9, cytochrome in isolated EVs and control cells (BRC and ADSC). (e) The western blotting demonstrating the presence of the SNAP25 protein common in neuronal cells, in small vesicles and abcence in small vesicles from ADSC only. (f) Immunocytochemistry quantification demonstrating the presence of the SNAP25 in ADSC (control), ADSC-CC, ADSC cultured with medium without EVs from BRC (Soluble factors-SF), and ASCD cultured Exo-BRC. (g) BRC expressing PalmtdTomato reporter (BRC + PalmtdTomato). (h-i) Culture of ADSC Exo-BRC + PalmtdTomato after day 3 and 7. (j) Co-cultured of ADSC with Exo-BRC + PalmtdTomato after day 3, images demonstrating the presence of EVs + PalmtdTomato together with ADSC. MERGE- merged image showing the extracellular vesicles communication between the cells in the co-culture. Bar = 20 μm. *p ≤ 0.05 and **p ≤ 0.001. The bars correspond to the means and the smaller bars are the standard error of the mean.