Isolation of Human Islet-Depleted Cells

Human islet-depleted cell fractions were obtained from organ donors deceased due to acute traumatic or anoxic death by Prodo Laboratories, Inc, and were shipped overnight to our laboratory (n = 21, see donor information in Supplementary Data 1). The flow sorting procedures were performed as previously described with some modifications^{14 (link)}. Briefly, exocrine tissue cells were incubated with FITC-conjugated UEA-1 (0.25 μg/ml, Vector Laboratories, Newark, CA, FL-1061-5) for 10 min. at 4 °C and washed with PBS. After washing, the cells were digested with TrypLE^TM Express (Life Technologies, Grand Island, NY, 12605-028) for 5−8 min at 37 °C. Cells were collected by centrifugation and washed with FACS buffer (10 mM EGTA, 2% FBS in PBS). After washing, cells were stained with Pacific blue-conjugated anti-CLA (BioLegend, San Diego, CA, 321308) and anti-7AAD (BioLegend, San Diego, CA, 420404) for 15 min at 4 °C. Cell pellets were collected by centrifugation and washed with PBS after staining. The cells were sorted using a FACSAria^TM II (BD Biosciences, San Diego, CA) and collected in 100% FBS. After sorting, cells were washed with serum-free Advanced DMEM/F-12 media (Life Technologies, Grand Island, NY, 12634-010).