GSM8996571: CAR72 manufactured CD19 CAR-T infusion product; Homo sapiens; RNA-Seq
Patient CAR T-cells were obtained by washes from commercial product infusion bags after patient infusion, and cryopreserved for batch analysis. Cells were washed with calcium and magnesium-free 1x PBS containing 0.04% BSA. After staining for viability using AO/PI and quantification with the Nexcelom Cellometer K2, cells were loaded onto the 10x Genomics Chromium Single Cell Controller at 1,000 cells/µl to encapsulate approximately 5,000 cells per sample. Inside nanoliter-sized Gelbeads in Emulsion (GEMs), single cells, reagents, and 10x Genomics gel beads underwent reverse transcription of poly-adenylated mRNA at 53°C. cDNA libraries were prepared using the 10x Genomics Chromium NextGEM Single Cell 5' Reagent Kit v2