GSM6084383: 14DMI; Mus musculus; RNA-Seq
Hearts were extracted from mice, cannulated, and perfused with PBS. They were then perfused with 5mL of collagenase type II solution diluted in PBS. They were then minced into small pieces using a razor blade and re-suspended in 10mL of collagenase solution on a rotator at 37 degrees celsius for 1 hour with frequent pipetting to ensure complete digestion. Cells were then passed through a 40 micron filter and centrifuged at 350g for 10 minutes to isolate the cardiac pericytes. Debris was removed using Debris Removal Kit and cells were re-suspended in FACS buffer. All cells were loaded to maximize the number of single cells acquired using the Chromium single Cell 3' Reagent Kit. Libraries were prepared according to manufacturer's instructions using the Chromium Single Cell 3' Library and Gel Bead Kit v.3 (10x Genomics). Libraries were sequenced on a NovaSeq Sequencer (Illumina) and with an average of 150,000,000 million reads sequenced per sample.