GSM9228770: Prostate cancer organoid cells, KBO114; Homo sapiens; RNA-Seq
Organoids were harvested from Matrigel and enzymatically dissociated into single cells using TrypLE™ Express Enzyme (1X) at 37°C for 10 minutes, with gentle pipetting. Cells were washed and resuspended in washing buffer (Advanced DMEM/F12, Glutamax 100X, HEPES solution 1M, Penicillin/Streptomycin X100). Cell viability>90% was confirmed for each PDO line by luciferase assay. Single-cell RNA sequencing (scRNA-seq) was performed using the Chromium Single Cell 3'  platform (10x Genomics, Pleasanton, CA, USA). Reverse transcription, cDNA amplification, and library construction were carried out using the Chromium Single Cell 3' Library & Gel Bead Kit v3 (10x Genomics) according to the manufacturer's protocol. Pooled libraries were sequenced on an Illumina NextSeq 2000 using a P3 flow cell with the following cycle configuration for 3' v3.1 Dual Index.