GSM3099849: Individual 7; Homo sapiens; RNA-Seq
Viable organoids were thawed and washed using DMEM, and digested with 0.05% trypsin (Corning 25-052-CI) containing DNase (Sigma Aldrich D4263-5VL) to generate single cell suspension. Cells were stained for FACS using fluorescently labeled antibodies for CD31 (eBiosciences 48-0319-42), CD45 (eBiosciences 48-9459-42), EpCAM (eBiosciences 50-9326-42), CD49f (eBiosciences 12-0495-82), SytoxBlue (Life Technologies S34857). We only proceeded with samples showing at least 80% viability as measured using SytoxBlue in FACS flow cytometry sorted cells were washed in PBS with 0.04% BSA and reseupended at a concentration of approximately 1000 cells/µl. Library generation for 10x Genomics v1 chemistry was performed following the Chromium Single Cell 3' Reagents Kits User Guide: CG00026 Rev B. Library generation for 10x Genomics v2 chemistry were performed following the Chromium Single Cell 3' Reagents Kits v2 User Guide: CG00052 Rev B.
GEO Accession
GSM3099849