Derivation of Injury-Responsive Dendritic Cells for Acute Brain Targeting and Therapeutic Protein Delivery in the Stroke-Injured Rat
Figure 4
In vitro characterization of protein cargo.
(A) Representative micrographs of harvested LV-hBDNF-transduced DCs. (B) Quantification of hBDNF protein in transduced-DC lysate (intracellular) and conditioned medium (secreted) by ELISA. Values reflect the combined average of 3 independent experiments, with each derived from separate lentiviral preps and DC culture preps. ** P<0.01, *** P<0.001 versus LV-GFP-transduced cultures by one-way ANOVA and Tukey post-hoc analysis. (C) Quantification of cortical neuron loss following OGD and DC treatment. Results were combined from at least 3 independent experiments. Error bars denote SEM. n = 46–60 wells/treatment group. * P<0.05, *** P<0.001 by one-way ANOVA and Tukey post-hoc analysis. (D) Representative fluorescent micrographs of LV-tBH4-transduced and LV-GFP-transduced-DC cultures. (E) Representative western blot showing anti-HisG immunoreactivity at 14 kDa for GFP-DC lysate or tBH4-DC lysate and conditioned medium. Time post-LV transduction is indicated for each sample. (F) Quantification of hippocampal neuron loss following glutamate exposure and DC treatment. Average neuron loss was calculated from at least 3 independent experiments. Error bars denote SEM. n = 24–48 wells/treatment group. * indicates significance by one-way ANOVA and Tukey post-hoc analysis with respect to each no insult control (vehicle or DC treatment alone, P<0.05). Scale bars: (A) and (D) 100 µm.