GSM8785345: hCD45+ cells, COMB7 CAR-M treatment; Homo sapiens; RNA-Seq
The tumor tissues were isolated from the mice and quickly immersed in PBS buffer. Samples were minced into about 0.5 mm3 pieces and enzymatically digested in digestion medium (PBS supplemented with collagenase I, collagenase II and collagenase IV).  The samples were incubated and digested at 37℃ for 60 min and then filtered using a 40 μm strainer. Cells were washed and resuspended in PBS buffer+0.1% FBS. Human CD45-positive cells were sorted by FACS. scRNA-seq library was constructed using the 10X Genomics Chromium system following manufacturer's instructions. In brief, the suspended cells were loaded into Chromium microfluidic chips and barcoded with a 10× Chromium Controller (10X Genomics). RNA from the barcoded cells was subsequently reverse-transcribed. The barcoded cDNA was purified and amplified, followed by fragmenting, A-tailing and ligation with adaptors.