title: GSM6760838: 50% WT GFP+ BMDMs and 50% WT BMDMs unstimulated; Mus musculus; RNA-Seq
library_construction_protocol: Bone marrow cells from femurs and tibia of wild-type C57BL/6N mice were cultured in Roswell Park Memorial Institute (RPMI) medium  supplemented with 10% heat-inactivated fetal bovine serum, 2 mM glutamine, 50 Î¼M 2-mercaptoethanol (Gibco), 100 U/ml penicillin, and 100 Î¼g/ml streptomycin (Gibco) and 50 ng/ml  of recombinant M-CSF (R&D Systems) on non-tissue culture treated 15 cm plates. On day 5 of culture, cells were harvested and 5 million cells were re-plated onto non-tissue culture  treated 10 cm plates in fresh media (same as that described above). After 2 additional days of culture, BMDMs from individual mice  were harvested by scraping. scRNAseq library constructions were carried out using the 10X Chromium Controller and the Chromium Next GEM Single Cell 3' Reagent Kits  (V3.1) in accordance with the manufacture's protocol. Prior to loading into the chromium controller, cells were washed once with PBS and resuspended in PBS at 700-1200 cells/ul for a target cell recovery of 7000 cells.