Primary adult rat mixed neural cell culture and in vitro hADSC-ex uptake

Mixed neural cell cultures were obtained through the prolonged culture of purified neurons obtained using OptiPrep™ density gradient (Sigma-Aldrich, D1556) separation. Primary adult rat cortical neurons were isolated from adult male Sprague-Dawley rats, as described previously [64]. Primary cortical neurons were cultured for 14-20 days (until glial cells appeared) to obtain mixed neural cells. To study the cellular uptake of hADSC-ex, we added DiI-hADSC-ex to the mixed neural cells and monitored the changes in DiI fluorescence intensity in each cell type for 24 h using a Cytation 5 Cell Imaging Multi-Mode Reader. The results were analyzed with Gen5 Data Analysis 3.0 Software.