GSM9081690: CD11b CD300e+/+, CD11b CD300e-/-; Mus musculus; RNA-Seq
Colonic adenomas and subcutaneous tumors were isolated and cut into small pieces of 2-4 mm in Petri dishes. The minced tissues were transferred into GentleMACS Tubes containing DMEM with 1 mg/mL collagenase IV (Sigma-Aldrich) and 0.5 mg/mL Dnase I. GentleMACS Tubes were attached onto a GentleMACS Octo Dissociator and tumors were maintained for 40 minutes at 37°C under continuous rotation. The resulting cell suspension was filtered through a 70 µm strainer and incubated with TruStain FcX™ PLUS. CD45+ cells from three CD300e+/+ and three CD300e-/- mice were isolated and independently labeled using Single Cell Labeling with the BD Single-Cell Multiplexing kit (BD Biosciences, #633793); next, the six samples were pooled prior FACS-sorting. CD11b+ myeloid cells were FACS-sorted as L/D-CD3e-CD19-CD11b+ cells. CD11b+ myeloid cells were processed using Single Cell Capture and cDNA Synthesis with the BD Rhapsody Express Single-Cell Analysis System. The resulting cDNA Libraries were prepared using mRNA Whole Transcriptome Analysis (WTA) and Sample Tag library preparation protocol.