GSM5257093: Myeloid cells from malignant pleural effusion_1; Mus musculus; RNA-Seq
After euthanization of the mice, all fluid was aspirated from the pleural cavity. The cell suspension was filtered through 100 μm and 40 μm cell strainers and centrifuged at 400 x g, 5 min at 4 ℃. Cells were resuspended in cell blood lysis buffer (0.15 M NH4Cl, 10 mM KHCO3, 0.1 mM EDTA) for 5 min, washed in PBS, resuspended in FACS-buffer (PBS, 2 mM EDTA, 25 mM HEPES) at a concentration of 1 million cells/100 μl and incubated for 20 min on ice with anti-CD45 andanti-CD11b antibodies. Labelled cells were washed with PBS and resuspended in FACS-buffer. FACS sorted alive CD45+ CD11b+ single cells were encapsulated in droplets using 10X Genomics GemCode Technology (Single Cell 3' v2 kit) and processed following manufacturer's specifications. Single Cell 3' v2