GSM4191570: Kit_tdTomato_aortic_graft_2wk; Mus musculus; RNA-Seq
Aoritc segments from donor BALB/c mice were transplanted into recipient Kit-CreERT2; ROSA26-tdTomato mice. Aortic grafts were harvested from Kit-CreERT2; ROSA26-tdTomato mice two or four weeks after surgury. Adventitial layer of the grafts was first dissected from the medial and neointimal layers. After three times washes of PBS, grafts were cut into fine pieces and digested in digestion solutions (which consist of 1 mg/ml papain, 0.5 mg/ml 1,4-Dithioerythritol, 0.156 mg/ml Taurine; 0.25 mg/ml bovine serum albumin in PBS) at 37 °C with reciprocal shaking for a total 40 min. Detached cells were then collected through a 40 μm cell strainer and suspended in DMEM with 10% FBS. Cells were further stained with DRAQ5 for 20 mins, followed by DAPI staining before loading to a Flow Cytometer. Single nucleated live tdTomato+ cells (DRAQ5+DAPI-tdTomato+) were sorted into PBS with 0.04% BSA and were then analyzed on a 10x Chromium instrument using Chromium Single Cell 3' Reagent Kit v3 chemistry (10X Genomics), following a standard protocol. Library was generated and sequenced on a Hiseq 2500 System (Illumina)
GEO Accession
GSM4191570