GSM8068078
Single cell suspensions of samples have been obtained by digesting the collagen hydrogel with 10'000 U/ml Collagenase Type 1 for 12 min at 37°C and by subsequently dissociating the cells with TrypLE Express for 10 min at 37°C. For multiplexing, cells were immunolabelled with TotalSeq-C anti-human Hashtag antibodies at a dilution of 1:500 in staining buffer (PBS+0.1% Bovine Serum Albumin) for 30 min on ice and washed 2 times in 4 ml staining buffer with intermittent centrifugation (300g, 3min). Cells were strained using tubes with 35μm cell strainer caps and counted and assessed for cell viability using an automated cell counter. 10X Chromium Next GEM Single Cell 5' v2 (Dual Index) chemistry (protocol CG000330 Rev F)