GSM8197687: WT_VD_1; Mus musculus; RNA-Seq
The aorta was washed three times in PBS, incubated at 37 °C for 40 minutes with an enzymatic dissociation cocktail (0.55 mg/mL collagenase I, 0.15 mg/mL collagenase IV and 0.07 mg/mL hyaluronidase in RPMI-1640 medium) and minced. The cell suspensions were filtered through 70 µm strainers and then pelleted by centrifugation at 400g for 5 minutes. The enzyme solution was then discarded and cells were resuspended in fresh PBS containing 0.04% BSA. Cell suspension was loaded onto the Chromium single cell controller (10x Genomics) to generate single-cell gel beads-in-emulsion (GEMs). Captured cells were lysed and the released RNA were barcoded through reverse transcription in individual GEMs. Reverse transcription was performed on a Thermal Cycler. Libraries were constructed using Single Cell 3' Library and Gel Bead Kit V3.1 according to the manufacturer's protocol.