Igg1 kappa

UEA1-FITC conjugated exocrine cell fraction was kept in suspension culture. At day 4, cell clusters were dissociated using StemPro™ Accutase™ Cell Dissociation Reagent (ThermoFisher Scientific, Waltham, Massachusetts, USA) following the protocol for dissociation of neurospheres, filtered over a 40 µm filter and cells were incubated with mouse monoclonal anti-human carbohydrate antigen 19.9 antibody (anti-CA19.9, Dako, Heverlee, Belgium; 2 µl per million cells in 200 µl) or isotype control (IgG1 kappa, Abcam, Cambridge, MA, USA; 2 µl per million cells in 200 µl) for 15 minutes at 4 °C. Cells were washed with FBS buffer (PBS + 3% FBS) and incubated for 15 minutes at 4 °C with secondary antibody Alexa fluor 647 anti-mouse (Jackson Laboratory, Westgrove, PA, USA; 2 µl per million cells in 200 µl). Analysis and cell sorting was performed on a BD FACSAria (BD Biosciences, Erembodegem, Belgium). Viable, single cells were gated based on forward and side scatter. After sort, cells were immediately collected in RLT buffer (Qiagen, Germantown, MD 20874, USA) and kept on ice. RNA extraction was immediately performed after sorting.