GSM9210596: Colon biopsy from Inactive UC patient, sample C; Homo sapiens; RNA-Seq
Colons were excised opened longitudinally, washed of fecal content with HBSS and cut into ~5mm pieces. Epithelium was separated from the LP by placing the tissue on a shaker for 15 minutes (150RPM, 37°C) using 10% FBS and 2mM EDTA HBSS solution. Remining LP tissue was minced and digested into single cell solution by shaking (200 RPM, 10-15 minutes) in collagenase solution (5% FBS, 1.5 mg/ml, collagenase type VIII and 40μg/ml DNase in HBSS), resulting cell suspensions were washed and filtered through a 40µm filter to ensure single cell separation.  Human colonic biopsies were obtained from consented inactive UC patients (Mayo endoscopic scores of 0) during routine screening and processed into single cell suspensions in a Liberase TH (0.1WU/mL), DNase I (0.5U/mL) in HBSS solution for 3 rounds of 10 min incubations at 37°C at 800RPM. Cell suspensions were washed and passed through a 40 strainer to ensure single cell separation. Libraries for both human and mouse samples were generated using Genomics Single Cell 3' Reagents kit (v3) in accordance with the manufacturer's protocols. Single cell suspensions were diluted in PBS 2%BSA targeting approximately 10,000 cells per sample. Single cells were sequenced to a read depth of ~ 20,000 reads per cell. Libraries were sequenced on an Illumina NovaSeq X Plus platform