characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-06-13T12:22:47Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-06-13T12:22:47Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA113413009
characteristics.characteristic: External Id
characteristics.text: Wellcome Sanger Institute
characteristics.characteristic: INSDC center name
characteristics.text: 2023-06-13T12:22:47Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-06-13T12:22:47Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS15408140
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12916:HCAHeartST13146207
characteristics.characteristic: Submitter Id
characteristics.text: 60 to 65
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: human
characteristics.characteristic: common name
characteristics.text: Protocols: Hearts were acquired after circulatory death (DCD) or after brain death (DBD). For DCD donors after confirmation of death there was a mandatory five-minute stand-off prior to sternotomy. In all cases the aorta was cross-clamped and cold cardioplegia solution was administered to the aortic root prior to cardiotomy. AV1, AV3, AV10, AV13 and AV14 were procured in the standard fashion and then immediately preserved with and transported on a hypothermic perfusion machine. AH2 was similarly preserved, but with immediate normothermic perfusion. It then underwent 4 hours of normothermic perfusion before samples were taken. For single nuclei sequencing, all donor samples were full-thickness myocardial tissues from the SAN, AVN, left and right atrium, left and right ventricles, interventricular septum and apex. Single nuclei were obtained from flash-frozen tissues using sectioning and mechanical homogenization. 5-10 mm thickness frozen tissues were first sectioned with cryostat in a 50 μm thickness section. All sections from each sample were homogenised using a 7 ml glass Dounce tissue grinder set (Merck) with 8–10 strokes of a loose pestle (A) and 8–10 strokes of a tight pestle (B) in homogenization buffer (250 mM sucrose, 25 mM KCl, 5 mM MgCl2, 10 mM Tris-HCl, 1 mM dithiothreitol (DTT), 1× protease inhibitor, 0.4 U μl−1 RNaseIn, 0.2 U μl−1 SUPERaseIn, 0.1% Triton X-100 in nuclease-free water). Homogenate was filtered through a 40-μm cell strainer (Corning). After centrifugation (500g, 5 min, 4 °C) the supernatant was removed and the pellet was resuspended in storage buffer (1× PBS, 4% bovine serum albumin (BSA), 0.2U μl−1 Protector RNaseIn). Nuclei were stained with 7-AAD Viability Staining Solution (BioLegend) and positive single nuclei were purified by fluorescent activated cell sorting (FACS) using MA900 Multi-Application Cell Sorter (Sony). Nuclei were further processed for 10x multiome paired RNA and ATACseq using the Chromium Controller (10X Genomics) according to the manufacturer’s protocol. cDNA library preparation was were carried out according to the manufacturer’s protocol.
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: normal
characteristics.characteristic: disease
characteristics.text: wild type genotype
characteristics.characteristic: genotype
characteristics.text: AV3
characteristics.characteristic: individual
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: sinoatrial node
characteristics.characteristic: organism part
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: Female
characteristics.characteristic: sex
characteristics.text: HCAHeartST13146207
characteristics.characteristic: title