characteristics.text: 2025-01-29
characteristics.characteristic: ENA first public
characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 1. CIC bioGUNE, BRTA 2. CIBERONC 3. Ikerbasque, Basque Foundation for Science 4. Traslational prostate cancer Research lab 5. Biochemistry and Molecular Biology Department. Spain.
characteristics.characteristic: INSDC center name
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS18102371
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-13764:AC1944_B
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: not collected
characteristics.characteristic: collection date
characteristics.text: human
characteristics.characteristic: common name
characteristics.text: Protocols: A benign prostate sample was collected adjacent to the tumor (contralateral lobe) from a patient with prostate cancer at the University Hospital of Basurto at the time of robotic prostatectomy following Ugalde-Olano et al., Mehods, 2015. The biopsy was kept in MACS medium, tissue storage solution until dissociation. The tissue was washed with cold phosphate-buffered saline (PBS; GIBCO) and minced into fragments of less than 0.4 mm. It was digested in filter-sterilized TH-liberase solution (2.5mg/ml; Sigma Aldrich) in complete medium composed of DMEM (GIBCO), Pen/strep (GIBCO), 1M MgCl2 (Sigma Aldrich), 10 mg/ml DNAse I (Sigma Aldrich) and Y-27632 dihydrochloride (10 μM; AbMole) at 37°C shaking at 800r.p.m for 40 minutes. The tissue solution was manually disaggregated every 10 min during incubation and centrifuged at 2400xg for 5 min. Subsequently, it was digested with filtered TrypLE (Life Technologies) + Y-27632 dihydrochloride under constant pipetting for 2 min. TrypLE inactivation was performed by adding FBS (GIBCO) and centrifuged at 2400xg for 5 min. The cell pellet was resuspended in PBS and filtered through a 40 μm filter and washed with PBS. The suspension was centrifuged at 2400xg for 5 minutes and resuspended in 200ul of PBS. Cells were counted and then centrifuged at 2400xg for 5 minutes and resuspended all in 10% DMSO, 50% FBS and 40% RPMI. Single cell library was prepared using “Chromium Next GEM Chip G Single Cell Kit”, 16 rxns PN-1000127, “Chromium Next GEM Single Cell 3ʹ Kit v3.1”, 4 rxns PN-1000269 and “Dual Index Kit TT Set A”, 96 rxns PN-1000215, following “Chromium Next GEM Single Cell 3 ’ Reagent Kits v3.1 (Dual Index) User Guide” (Document number CG000315). Single cell library was prepared using “Chromium Next GEM Chip G Single Cell Kit”, 16 rxns PN-1000127, “Chromium Next GEM Single Cell 3ʹ Kit v3.1”, 4 rxns PN-1000269 and “Dual Index Kit TT Set A”, 96 rxns PN-1000215, following “Chromium Next GEM Single Cell 3 ’ Reagent Kits v3.1 (Dual Index) User Guide” (Document number CG000315).
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: prostate cancer
characteristics.characteristic: disease
characteristics.text: not collected
characteristics.characteristic: geographic location (country and/or sea)
characteristics.text: not applicable
characteristics.characteristic: isolate
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: prostate gland
characteristics.characteristic: organism part
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: male
characteristics.characteristic: sex
characteristics.text: AC1944_B
characteristics.characteristic: title