GSM4779215: Facs1; Mus musculus; RNA-Seq
Mammary glands number 4 were collected and pooled from a total of four 10-week old, female FVB/NJ mice. Glands were minced into pieces ~1mm in diameter and were incubated with a 2mg/ml collagenase type IV solution at 37C while shaking for 1 hour. Digested organoids were collected by differential centrifugation. Collected organoids were further dissociated with trypsin into single cells. Cells were stained for flow cytometry using fluorescently labeled antibodies for CD49f, EpCAM, CD31, CD45, Ter119, and SytoxBlue. Live epithelial cells were collected for sequencing Library generation for 10x Genomics v2 chemistry was performed following the Chromium Single Cell 3' Reagents Kits v2 User Guide: CG00052 Rev B. Libraries were sequenced on the Illumina HiSeq4000 platform targeting approximately 50,000 reads per cells.