characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2022-05-01T00:14:21Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2022-05-01T00:14:21Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA13173515
characteristics.characteristic: External Id
characteristics.text: Section of Genetics and Genomics, Department of Metabolism, Digestion and Reproduction, Imperial College London
characteristics.characteristic: INSDC center name
characteristics.text: 2022-05-01T00:14:21Z
characteristics.characteristic: INSDC first public
characteristics.text: 2022-05-01T00:14:21Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS10777073
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-11477:Haster_Control3
characteristics.characteristic: Submitter Id
characteristics.text: 7
characteristics.unit: week
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: To isolate pancreatic islets, the pancreas was inflated with an ice-cold collagenase P solution (1 mg/mL in Hanks' balanced salt solution (HBSS) buffer, Roche) injected through the main pancreatic duct. The pancreas was then dissected and incubated at 37ºC for 8 min with continuous agitation. The pancreatic tissue was disaggregated by gentle suction through a needle, and cell suspension was washed 4 times with cold HBSS-0.5% BSA. The islet pellet was resuspended in 7 mL Histopaque containing a 7:3 ratio of pre-cooled Histopaque 1077 (Merck) and Histopaque 1119 (Merck), and 7 mL of HBSS-0.5% BSA was layered on top of the Histopaque. The gradient was centrifuged at 950 g for 20 mins at RT. The interphase containing the islets was collected, washed 3 times with HBSS-0.5% BSA and islets were finally further enriched by gentle aspiration with a pipette under a stereomicroscope. Islets were allowed to recover for 2 days in culture in RPMI containing 11 mM glucose supplemented with 10% fetal calf serum and penicillin-streptomycin (1:100; Invitrogen) at 37°C and 5% CO2. Cultured mouse islets were dissociated with Accutase (Merk) for 15 min at 37ºC. Islet cell suspensions were centrifuged at 600g 3 min and resuspended in cell culture medium with DAPI before FACS sorting to remove dead cells and doublets. After sorting, cells were centrifuged at 600g for 3 min and resuspended in PBS/0.04% BSA. Nucleic acid were extracted following the manufacturer’s instructions for 10x Genomics Chromium Single Cell 3′ Reagent Kit v3 Libraries were prepared following the manufacturer’s instructions for 10x Genomics Chromium Single Cell 3′ Reagent Kit v3
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: Haster +/+;Pdx1-Cre
characteristics.characteristic: genotype
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: islet of Langerhans
characteristics.characteristic: organism part
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: female
characteristics.characteristic: sex
characteristics.text: C57BL/6
characteristics.characteristic: strain
characteristics.text: Haster_Control3
characteristics.characteristic: title