Figure 6.

Neuroprotective properties of AD-MSCs exosome content. Assessment of neurite outgrowth (A,B) in human nociceptors grown as neurospheres and exposed for 5 days to either NGF, sAAT, and dAAT exosomes (10 µg/mL) or the total secretome. Phase contrast images of the spheres were taken (A) and neurite length was measured on day 1 and 5 post treatment (B). Scale bar, 150 µm. Restoration of neurite outgrowth (C,D) in human sensory neurons dissociated as single cells, and re-plated and exposed for 5 days to either NGF, sAAT, and dAAT exosomes (10 µg/mL) or the total secretome. (C) Representative pictures of the neuronal network were taken on day 1 and 5 and the network properties were quantified using a customized version of the Angiogenesis Analyzer ImageJ plugin. Scale bar, 100 µm. (a) Initial image sample of NGF-treated human sensory neurons. (b) Network analysis with customized version of «Angiogenesis Analyzer» and vectorial objects as defined in (A), and (c,d) neuronal network developed in response to sAAT and dAAT exosomes, respectively. (D) Quantification of the total branching length developed and normalized to the analyzed area. * shows significance versus the control group (NGF) on day 1 and * on day 5, * p < 0.05, ** p < 0.01, and *** p < 0.001. (E) Cytokine antibody array membranes of sAAT and dAAT exosome content with positive (blue square) and negative (yellow squares) internal controls. An equal amount of exosomal proteins was loaded for each membrane. (F) Quantification of the antibody array using ImageJ software.