GSM6740282: Donor 3 - 24 hours treatment; Homo sapiens; RNA-Seq
After treatment islets were dispersed into single cells by dynamic incubation with 0.025% trypsin (Gibco) supplemented with 10 mg/mL DNase (Pulmozyme, Genentech) for 6–7 min and filtered through a 40 μm cell-strainer to remove undigested material. Islets exposed to the different treatments were multiplexed per timepoint and donor using cell hashing. Briefly, each treated islet cell suspension was incubated with a pool of barcoded antibodies directed against B2M and CD298 (TotalSeq-A BioLegend). Labeled cell suspensions were pooled per donor and time point and processed for single-cell mRNA sequencing. Prior to loading the 10x Chromium controller, the pooled cell suspensions were counted to assess cell integrity and concentration. Cell suspensions were loaded, and the sequencing libraries were prepared following the standard 10x Genomics 3' V3 chemistry protocol.