SAMN31872531
External Id
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences
INSDC center name
2023-12-31T01:09:08.793Z
INSDC first public
2023-12-31T01:09:08.793Z
INSDC last update
SRS15864341
INSDC secondary accession
live
INSDC status
Human
NCBI submission model
Human.1.0
NCBI submission package
SRS15864341
SRA accession
not applicable
age
not specified
biomaterial_provider
HepaRG
cell_line
immortalized hepatic cell line
cell_type
*** Cell culture *** Cells were grown in a 5% CO2 humidified atmosphere at 37°C. HepaRG and HepaRGNTCP cells were cultured in William’s E medium containing GlutaMAX, 5 µg/ml human insulin, 50 µM hydrocortisone hemisuccinate, 100 IU/ml penicillin, 100 µg/ml streptomycin, and 10% fetal bovine serum (Fetal clone II). HepaRGNTCP cell culture medium was supplemented with 5 µg/mL blasticidin and 100 µM zeocin. Medium was renewed wice a week. Cells were split once a week till passage 16-18. For HDV infection HepaRG cells were seeded at the density allowing cells to be sub-confluent the next day. After two weeks of culture, the medium was supplemented with 1.8% DMSO, and the cells were left for two more weeks with the medium change twice a week. *** HDV infection *** 4 weeks post-seeding cells were infected with HDV at MOI 10 in the presence of DMSO and 4% PEG 8000 (stock solution 40% PEG 8000 in PBS, sterile-filtered) for 24h. Mock cells were treated only with PEG and DMSO. Then cells were washed with PBS and incubated in the medium with DMSO. HDV replication reaches the maximum on day 4 and is stable till day 8-9, then starts to decline. In this study 6-7 days infections were performed. HepaRGNTCP cells were seeded at the density allowing cells to be sub-confluent the next day. After 4-5 days of culture the medium was supplemented with 1.8% DMSO and 1 µg/ml tetracycline. HDV infection was done on day 7-8 post seeding equally to HepaRG cell line. All infections were performed in duplicates or triplicates. *** treatment *** Ruxolitinib was given to the treated samples at concentration 1 µM for 4 days. *** RNA extraction *** HepaRGNTCP cells were seeded in 6-well plates and infected with HDV for 7 days as described above. Cells were washed with PBS, harvested by scrapping and precipitated. The precipitates were stored in RNAlater solution at -80°C until further use. Total RNA was extracted from cell samples using Qiagen RNeasy Plus Mini kit following manufacturer’s instructions (Qiagen, Hilden, Germany).
description
not applicable
isolate
Homo sapiens
http://purl.obolibrary.org/obo/NCBITaxon_9606
organism
replicate 2
replicate
not applicable
sex
liver
tissue
HepaRG cells (non-infected, non-treated) replicate 2
title
none
treatment