characteristics.text: 2024-01-01
characteristics.characteristic: ENA first public
characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: SAMEA112942597
characteristics.characteristic: External Id
characteristics.text: Odense University Hospital/University of Southern Denmark
characteristics.characteristic: INSDC center name
characteristics.text: 2023-04-20T10:32:17Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS14943179
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12865:Sample 2
characteristics.characteristic: Submitter Id
characteristics.text: 69
characteristics.unit: year
characteristics.characteristic: age
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: adipose-derived regenerative cell
characteristics.characteristic: cell_type
characteristics.text: human
characteristics.characteristic: common name
characteristics.text: Protocols: ADRCs were isolated from human lipoaspirates using an automated processing Celution® 800/IV device (Lorem Cytori Therapeutics) and used immediately for further enrichment of CD31-postitive ADRCs. ADRCs were subjected to red cell lysis before enrichment of CD31-postitive ADRC subpopulations was accomplished by magnetic cell sorting (MACS) using MS separation columns and CD31 microbead kits (Miltenyi Biotech). To enable single-cell RNA sequencing (scRNA-seq), at least 1,000,000 CD31+ ADRCs from each of four subjects (one male and three females) were methanol-fixed and stored at -80°C. Briefly, the cells were resuspended in 500 µl PBS with 1% BSA and 1 U/µl RNAsin PLUS RNase Inhibitor (Promega) and incubated on ice for 5 min to ensure the integrity of the RNA in the cells. The cells were passed through a 40 µm Flowmi® Cell strainer (VWR) to create a single-cell suspension. The cells were centrifuged at 400g for 3 min at 4°C, and washed in 1 ml RNase-free PBS and subsequently centrifuged at 400g for 3 min at 4°C, before the cell pellet was resuspended in 200 µl RNase-free PBS and 800 µl ice-cold methanol gently added dropwise to the cells. The cells were fixed for 30 min at -20°C and subsequently stored at -80°C until use. Before sequencing, fixed cells were thawed and rehydrated. Briefly, the cells were centrifuged at 1,000g for 10 min at 4°C and the supernatant was removed. The cell pellet was resuspended (1,000 cells/µl) in a rehydration buffer (3x saline sodium citrate, 0.04% BSA, 1 U/µl RNAsin Plus, 40 mM Dithiothreitol) and incubated at 4°C for 30 min. Immediately following rehydration, 8,000 cells were loaded onto the 10x Genomics Chromium controller (10X Genomics). Libraries were prepared according to the instructions of the manufacturer using the 10x Genomics Single-Cell 3’ v3, Chromium Single Cell B Chip Kit, 48 runs (10X Genomics).
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: CD31-positive
characteristics.characteristic: immunophenotype
characteristics.text: Pt 2
characteristics.characteristic: individual
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: subcutaneous adipose tissue
characteristics.characteristic: organism part
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: female
characteristics.characteristic: sex
characteristics.text: Sample 2
characteristics.characteristic: title