characteristics.text: 2025-12-01
characteristics.characteristic: ENA first public
characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: Latvian Biomedical research and study centre
characteristics.characteristic: INSDC center name
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS22551435
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-14667:Hs68-EV1
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: Hs68
characteristics.characteristic: cell_line
characteristics.text: fibroblast of dermis
characteristics.characteristic: cell_type
characteristics.text: not collected
characteristics.characteristic: collection date
characteristics.text: human
characteristics.characteristic: common name
characteristics.text: Protocols: Collected SW620 bioreactor culture supernatants were pooled and centrifuged at 3,000 × g for 30 minutes at 4°C to remove cellular debris, followed by filtration through 0.2 µm syringe filters. The filtered supernatants were concentrated to ≤10 mL using 100 kDa Amicon Ultra-15 filter units (Merck Millipore, Germany) at 3,220 × g, 4°C. Concentrates were further centrifuged at 10,000 × g for 10 minutes at 4°C to remove residual debris. Plasma samples were thawed at 25°C, vortexed, and centrifuged at 10,000 × g for 10 minutes.  Supernatants or plasma (10 or 1 mL, respectively) were loaded onto pre-equilibrated Izon qEV10 or qEV1 35 nm GEN2 size exclusion chromatography (SEC) columns (Izon, New Zealand), respectively, and EVs were eluted with DEPC-treated PBS (Fisher BioReagents, USA) following the manufacturers protocol. EV-enriched fractions were identified by dynamic light scattering on a Zetasizer Nano ZS instrument (Malvern Panalytical, UK). Fractions where >70% of particles exceeded 30 nm in size were pooled as EV fractions and following smaller particle fractions were pooled as protein fractions (PR), concentrated to 100 µL using 3 kDa Amicon Ultra-2 filters (Merck Millipore, Germany) at 7,500 × g, 4°C, and stored at −80°C. HS68, WI-38, and SW480 cells were seeded into 12-well plates at a density of 1 × 10⁵ cells/well in DMEM/F12 medium supplemented with 10% CDM-HD, 1× Antibiotic-Antimycotic, and 2 mM L-glutamine. EVs were added to the cells at a concentration of 1 × 10⁵ EVs/cell. 10 µg of proteins was added to the PF wells and negative control wells received 100 µL of DEPC-treated PBS.  After a 24-hour incubation at 37°C with 5% CO₂ and 100% humidity, cells were washed with PBS and lysed in 350 µL RLT buffer containing 1% β-mercaptoethanol (RNeasy Mini Kit, Qiagen, Germany). Lysates were vortexed for 30 seconds and stored at −80°C. RNA was extracted using the RNeasy Mini Kit with additional on-column DNase digestion according to the manufacturer’s protocol. RNA was eluted in 40 µL of nuclease-free water and stored at −80°C. RNA concentration was determined using a NanoDrop ND-1000 spectrophotometer (Thermo Fisher Scientific, USA). Total RNA (200 ng) was treated with the MGIEasy rRNA Depletion Kit (MGI, China) to remove ribosomal RNA, followed by library construction using the MGIEasy RNA Directional Library Prep Set (MGI, China). Libraries were prepared by fragmenting RNA to ~250 bp, synthesizing cDNA, performing end-repair and A-tailing, adding unique adapters for sample identification, and amplifying the library via PCR. Library quality was assessed using Qubit dsDNA High Sensitivity Assay Kit (Thermo Fisher Scientific, USA) and Agilent High Sensitivity DNA Chip (Agilent, USA). Libraries were pooled at 1 pmol, denatured, circularized, and quantified with the Qubit ssDNA Assay Kit (Thermo Fisher Scientific, USA).
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: Aspartoacylase Deficiency
characteristics.characteristic: disease
characteristics.text: wild type genotype
characteristics.characteristic: genotype
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: dermis
characteristics.characteristic: organism part
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: EV
characteristics.characteristic: stimulus
characteristics.text: Hs68-EV1
characteristics.characteristic: title