GSM7916355
PWD5 wound tissue was dissected from the surrounding edge and healthy tissue, minced with scissors, and then dissociated with 10 ml solution of 2.7 mg/mL collagenase P (Sigma), 1mM pyruvate, 10 mM HEPES, in RPMI basal media at 37°C for 2 hours at 37C, triturating every 15 minutes. Digestion was halted with the addition of 2ml 2% FBS and live cells were isolated using Miltenyi dead cell separation kit and MS columns, per manufacturer's instructions. Cells were finally suspended in 1ml 0.04% BSA and kept on ice until processed for library preparation. RNA library was prepared using a 10X Chromium V3.1 kit and sequenced with NovaSeq on an S4 flow cell (UCI Genomic High Throughput Facility).