AC1944_B
Homo sapiens
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).
ENA first public
2025-01-29
INSDC center name
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.
INSDC status
public
Submitter Id
E-MTAB-13764:AC1944_B
broker name
ArrayExpress
collection date
not collected
common name
human
developmental stage
adult
disease
prostate cancer
geographic location (country and/or sea)
not collected
isolate
not applicable
organism part
prostate gland
sample name
E-MTAB-13764:AC1944_B
scientific_name
Homo sapiens
sex
male