Isolation of Placental Amnion-Derived MSCs

Primary culture of MSCs was obtained from placental amnion by enzymatic method [25 (link)]. Amnion membranes were washed with PBS with 10% Ciprofloxacin (Fresenius Kabi, Bad Homburg, Germany), then dissected into small pieces and incubated in presence of 0.25% trypsin for 1 hour at 37°C. Following trypsin digestion, samples were filtered through 100 μm cell strainer (BD Biosciences, Durhan, USA), the cell suspension was centrifuged for 5 min at 1200 rpm (Heraeus Multifuge 1S-R, Thermo Fisher Scientific GmbH, Dreieich, Germany), the cell pellet was resuspended in MSC growth medium and plated into 10 cm cell dishes (Cellstar, Greiner BioOne, Frickenhausen, Germany).