{"EMPIAR-13172":{"imagesets":[{"segmentations":[],"name":"Extracellular vesicles (EVs) isolated from PANC-1, PPCL-68, and hTERT-HPNE cell lines conditioned media","directory":"data","category":"micrographs - single frame","header_format":"TIFF","data_format":"TIFF","num_images_or_tilt_series":3,"frames_per_image":1,"frame_range_min":null,"frame_range_max":null,"voxel_type":"UNSIGNED 16 BIT INTEGER","pixel_width":3.82,"pixel_height":3.82,"micrographs_file_pattern":"","picked_particles_file_pattern":"","picked_particles_directory":"","details":"EV samples (∼3.5 μL) were loaded onto glow-discharged, perforated carbon-coated grids (2/1-3C C-Flat; Protochips, Raleigh, NC), blotted with filter paper, and rapidly plunged into liquid ethane. These grids were stored in liquid nitrogen. Before imaging, the grids were placed on cryo-specimen holder (626 cryo-specimen holder, Gatan, Warrendale, PA) and maintained at -180°C. The digital micrographs were captured using a Tecnai F20 Twin transmission electron microscope (ThermoFisher Scientific, Hillsboro, OR) equipped with a Gatan US4000 CCD or a Teitz XF416 camera.","image_width":"4096","image_height":"4096"}],"workflow_file":null,"grant_references":[],"version_history":[],"title":"Cryogenic transmission electron microscopy images of extracellular vesicles isolated from conditioned media of pancreatic cancer and normal pancreatic epithelial cell lines","principal_investigator":[{"author_orcid":"0000-0003-4877-7583","middle_name":"K","organization":"Department of Oncology, Georgetown University","street":"3970 Reservoir Road NW","town_or_city":"Washington","state_or_province":"District of Colombia","post_or_zip":"20007","telephone":null,"fax":null,"first_name":"Amrita","last_name":"Cheema","email":"akc27 [at] georgetown.edu","country":"United States","entry":"EMPIAR-13172"}],"status":"REL","deposition_date":"2025-12-13","release_date":"2025-12-24","obsolete_date":null,"update_date":"2025-12-24","corresponding_author":{"author":{"author_orcid":"0000-0003-4877-7583","middle_name":"K","organization":"Department of Oncology, Georgetown University","street":"3970 Reservoir Road NW","town_or_city":"Washington","state_or_province":"District of Colombia","post_or_zip":"20007","first_name":"Amrita","last_name":"Cheema","country":"United States"}},"authors":[{"author":{"name":"Singh BS","author_orcid":"0009-0000-9461-6110"}},{"author":{"name":"Gaur PG","author_orcid":"0000-0002-9718-3565"}},{"author":{"name":"Bose PB","author_orcid":"0000-0001-9751-3299"}},{"author":{"name":"Zhang YJ","author_orcid":"0009-0002-8531-6604"}},{"author":{"name":"Li YL","author_orcid":"0000-0001-9200-1016"}},{"author":{"name":"Zhang ZZ","author_orcid":null}},{"author":{"name":"Kandhavelu JK","author_orcid":"0000-0003-3511-3040"}},{"author":{"name":"Klotzbier WK","author_orcid":null}},{"author":{"name":"Jayatilake MJ","author_orcid":"0000-0002-5780-9391"}},{"author":{"name":"Bansal SB","author_orcid":"0000-0002-5095-5687"}},{"author":{"name":"Farhan MF","author_orcid":null}},{"author":{"name":"Deol SD","author_orcid":"0009-0006-8410-9928"}},{"author":{"name":"Banerjee PPB","author_orcid":null}},{"author":{"name":"Unger KU","author_orcid":null}},{"author":{"name":"Gupta SG","author_orcid":"0000-0003-0106-595X"}},{"author":{"name":"Verma VV","author_orcid":"0000-0001-8129-4140"}},{"author":{"name":"Cheema AKC","author_orcid":"0000-0003-4877-7583"}}],"cross_references":[],"biostudies_references":[],"idr_references":[],"empiar_references":[],"citation":[{"authors":[{"name":"Singh BS","author_orcid":"0009-0000-9461-6110"},{"name":"Gaur PG","author_orcid":"0000-0002-9718-3565"},{"name":"Bose PB","author_orcid":"0000-0001-9751-3299"},{"name":"Zhang YJ","author_orcid":"0009-0002-8531-6604"},{"name":"Li YL","author_orcid":"0000-0001-9200-1016"},{"name":"Zhang ZZ","author_orcid":"0009-0001-3254-3494"},{"name":"Kandhavelu JK","author_orcid":"0000-0003-3511-3040"},{"name":"Klotzbier WK","author_orcid":null},{"name":"Jayatilake MJ","author_orcid":"0000-0002-5780-9391"},{"name":"Bansal SB","author_orcid":"0000-0002-5095-5687"},{"name":"Farhan MF","author_orcid":null},{"name":"Deol SD","author_orcid":"0009-0006-8410-9928"},{"name":"Banerjee PPB","author_orcid":null},{"name":"Unger KU","author_orcid":null},{"name":"Gupta SG","author_orcid":"0000-0003-0106-595X"},{"name":"Verma VV","author_orcid":"0000-0001-8129-4140"},{"name":"Cheema AKC","author_orcid":"0000-0003-4877-7583"}],"editors":[],"published":false,"j_or_nj_citation":true,"title":"Extracellular vesicle derived miRNA-182-5p educates macrophages towards an immunosuppressive phenotype in pancreatic cancer","volume":null,"country":"United States","first_page":null,"last_page":null,"year":null,"language":"English","doi":null,"pubmedid":null,"details":"EV isolation and Characterization\nEVs were isolated from the conditioned media using size exclusion chromatography (SEC). When the cells reached 50-60% confluency, the media was discarded and the cells were washed with PBS. The media for each cancer cell line was replenished with their respective basal media containing 10% (v/v) exosome-depleted FBS (Gibco, A2720801) and 1% (v/v) Penicillin-Streptomycin (Gibco, #15140122). For hTERT-HPNE cells, the medium was replaced with fresh serum-free medium (K-SFM) supplemented with bovine pituitary extract, human recombinant epidermal growth factor (Gibco, #37010022), and 1% (v/v) Penicillin-Streptomycin (Gibco, #15140122). Cells were grown for 48 h. Subsequently, the media was collected and centrifuged twice at 2500 x g for 10 mins at 25°C to remove any debris or cells. The supernatants were then concentrated using 100 kDa Centricon Plus-70 filters (Millipore, #UFC710008) by spinning at 3500 x g for 20 mins at 25°C. Filtered flow-through was discarded, and the concentrate was collected as per the manufacturer’s instructions. The process was repeated until all the supernatant was concentrated. The EVs were isolated from the concentrate via SEC by utilizing a qEV2/70 nm column (Izon Science, IC270) with an automated fraction collector, according to the manufacturer’s protocol. The first three fractions were pooled, lyophilized, and resuspended in DPBS and stored in -80°C until further characterization and experiments.\n\nCryogenic transmission electron microscopy (cryoTEM)\nEV samples (∼3.5 μL) were loaded onto glow-discharged, perforated carbon-coated grids (2/1-3C C-Flat; Protochips, Raleigh, NC), blotted with filter paper, and rapidly plunged into liquid ethane. These grids were stored in liquid nitrogen. Before imaging, the grids were placed on cryo-specimen holder (626 cryo-specimen holder, Gatan, Warrendale, PA) and maintained at -180°C. The digital micrographs were captured using a Tecnai F20 Twin transmission electron microscope (ThermoFisher Scientific, Hillsboro, OR) equipped with a Gatan US4000 CCD or a Teitz XF416 camera.","book_chapter_title":null,"publisher":null,"publication_location":null,"journal":"Signal Transduction and Targeted Therapy","journal_abbreviation":"Sig Transduct Target Ther","issue":null,"preprint":false}],"dataset_size":"96.1 MB","experiment_type":"SBF-SEM","scale":"cell","entry_doi":"10.6019/EMPIAR-13172"}}