characteristics: ERC000011
characteristics: ENA-CHECKLIST
characteristics: 2022-09-30T00:19:51Z
characteristics: ENA-FIRST-PUBLIC
characteristics: 2022-09-30T00:19:51Z
characteristics: ENA-LAST-UPDATE
characteristics: SAMEA10273810
characteristics: External Id
characteristics: Latvian Biomedical Research and Study Centre
characteristics: INSDC center name
characteristics: 2022-09-30T00:19:51Z
characteristics: INSDC first public
characteristics: 2022-09-30T00:19:51Z
characteristics: INSDC last update
characteristics: public
characteristics: INSDC status
characteristics: ERS7928667
characteristics: SRA accession
characteristics: E-MTAB-11022:Hs68-DB1202
characteristics: Submitter Id
characteristics: ArrayExpress
characteristics: broker name
characteristics: Hs68
characteristics: cell line
characteristics: fibroblast
characteristics: cell type
characteristics: human
characteristics: common name
characteristics: Protocols: Cells were seeded in DMEM-F12 complete medium at a density of 1x105 cells per well in 12-well plates and 1x104 EVs per cell (total 1x109 EVs / well) were added to the cells and they were grown as a monolayer for 48 hours. Untreated control cells were grown in parallel. After that, the cells were washed with PBS and lysed with 1ml TRI-Reagent® (Sigma Aldrich, USA) and RNA was extracted according to manufacturer’s protocol. The cells were washed with PBS and lysed with 1ml TRI-Reagent® (Sigma Aldrich, USA) and RNA was extracted according to manufacturer’s protocol. Briefly, 200 μl of chloroform was added, the samples were centrifuged at 12 000g for 15 min at +4˚C, the aqueous phase was transferred to a new tube and isopropanol was used to precipitate the RNA by centrifuging at 12 000g for 10 min at +4˚C. After that, the RNA was washed with 70% Ethanol-DEPC and the DNA was removed with DNA-free™ (Thermo Fisher Scientific, USA) DNA removal kit according to manufacturer's protocol. The RNA concentration was measured with NanoDrop 1000 (Thermo Ficher Scientific, USA) and the integrity of the RNA was measured with Agilent Bioanalyzer using RNA Pico chip (Agilent Technologies, USA). For transcriptome sequencing, the rRNA was removed from 200 ng of total RNA using MGIEasy rRNA depletion kit (MGI, China). The transcriptome libraries were built using MGIEasy RNA Directional Library Prep Kit (MGI, China) according to manufacturer’s protocol. Briefly, the RNA was fragmented into 250 bp pieces, and it was reverse transcribed, and the second strand was synthesized. Adapters were ligated according to the manufacturer’s recommendations and the product was amplified with PCR.
characteristics: description
characteristics: adult
characteristics: developmental stage
characteristics: normal
characteristics: disease
characteristics: Homo sapiens
characteristics: organism
characteristics: Homo sapiens
characteristics: organism
characteristics: foreskin
characteristics: organism part
characteristics: Homo sapiens
characteristics: scientific_name
characteristics: male
characteristics: sex
characteristics: extracellular vesicles from urine of healthy male
characteristics: stimulus
characteristics: Hs68-DB1202
characteristics: title