GSM8140025: E13.5TAM_E16.5_42; Mus musculus; RNA-Seq
For the isolation of adult mouse islets, firstly, euthanize mice and expose abdominal organs. Next, clamp the duodenum at the upper and lower position where the common bile duct enters the duodenum. Then, inject pre-cold 1 mg/ml collagenase P into the common bile duct to inflate whole pancreas. Digest pancreas in 37℃ water bath for 6-8 min with gently shaking, and then manually pick out islets using microscope. These islets were digested into single-cell suspension with 0.25% trypsin for 5 min at 37℃. For the digestion of embryonic mouse pancreas, firstly, E14.5-18.5 pancreas were dissected and digested with 1 mg/ml collagenase P for 3-5 min which varies depending on pancreas size. Then, dissociated pancreas were treated with 0.25% trypsin for 5 min at 37℃. Cell suspension obtained after digestion can be directly sorted using the endogenous fluorescent protein carried by the cells and BD FACSAria Fusion flow cytometer. For smart-seq3xpress, single cells were collected according to two strategies: one was to sort single cells into 384-well plates containing lysis buffer, and the other was to pick single cells manually from FACS-purified bulk cells by using a mouth pipette.