GSM6622111: Young; Mus musculus; RNA-Seq
For the young and old pituitary sample, anesthetized mice were perfused via the aorta with cold PBS and dissected pituitary was placed into an enzyme mix containing 0.5% w/v collagenase type 2, 0.25% trypsin, in Hanks balanced salt solution for 2-3 hours at 37°C. The tissue and cells were gently triturated and pelleted at 900g for 3 minutes, and resuspended for further single cell sorting. All live cells from young or old pituitary negative for DAPI  staining were sorted for further scRNA-seq. For the Igha-Cre sample, zsGreen positive and DAPI negative pituitary cells were sorted for sequencing. For the Epcam sample, old (18-24M) pituitary cells were stained with DAPI and FITC conjugated Epcam antibody, then DAPI negative and Epcam positive cells were sorted for further processing. Libraries were prepared for sequencing using standard Illumina protocols