GSM8879328: CbO+NSC73.5; Homo sapiens; RNA-Seq
Pooled samples of 5 CbO were taken at day 7 and day 35, as well as CbO-MB at day 49. First, organoids were dissociated using the Neural dissociation kit (MACS Miltenyi Biotec) into single cell suspension. Subsequent debris removal was then performed using Debris removal solution according to manufacturer protocol (Miltenyi Biotec, #130-109-398) to obtain a clean sample of highly viable cells. Cells were resuspended in loading buffer (0.04% BSA in PBS), strained through a 70 μm cell strainer and counted using a hemacytometer with trypan blue staining. Library preparation was performed at the Single Cell Genomics Facility (UCL, Cancer Institute) using the Chromium Next GEM Single Cell 3' Kit v3.1 (10X Genomics, 1000268) along with the Chromium Next GEM Chip G Single Cell Kit (10X Genomics, 1000120) according to the manufacturer's instructions. Pooled samples libraries were sequenced with lane sequencing on Novaseq 6000 S4 (Novogene).