Single-cell analysis of Villin-Cre;Apc min;Lef1 fl/fl (VApcMinL) and ApcMin intestinal adenoma cells at the age of 11 wks

Comparison of Villin-Cre;Apc min;Lef1 fl/fl (VApcMinL) and ApcMin adenoma cells at the age of 11 wks. 2-3 male mice and 3-10 tumors per group were used in the experiment. Experiment was repeated twice (ApcMin2 and VApcMinL2 samples). We analyzed two samples, aiming with both samples to 5,000-8,000 cells. For tumor dissociation, 3-10 tumors of approximately same size and from similar locations in the intestines were removed from 11-week-old ApcMin and VApcMinL mice. Tumors were incubated with enzymes (4 mg/ml Dispase II, 1 mg/ml Collagenase I, 1 mg/ml Collagenase H and 1 U/ml DNase I) for 30 min at +32°C, followed by incubation with trypsin-EDTA for 15 min at +37°C. Cells were filtered through 40 μm mesh and subjected to scRNA-seq without FACS sorting.