Sample 10
Mus musculus domesticus
Protocols: Primary hepatocytes were isolated by ex vivo perfusion of liver left lateral lobes using the gentleMACS Octo Dissociator with Heaters and the associated Liver Perfusion Kit. Cells were cultured at 37°C and 5% CO2 for 3 h in collagen-coated 6-well plates containing DMEM supplemented with 5% FCS and 1% Pen Strep (Charni-Natan and Goldstein 2020), cultured overnight (for 14-21 h) in William's E medium supplemented with 2 mM GlutaMAX and 1% Pen Strep (Charni-Natan and Goldstein 2020), cultured for 30 min in William's E medium supplemented with 2 mM GlutaMAX (Charni-Natan and Goldstein 2020). siRNAs were delivered using Lipofectamine RNAiMAX Reagent according to the manufacturer's instructions (25 pmol siRNA and 7.5µl RNAiMAX per well). At 48 hpt, primary hepatocytes were incubated in 0.25% Trypsin-EDTA solution for 6 min at 37°C and 5% CO2, transferred to 2 ml screw cap micro tubes, rinsed once with PBS to dilute out residual proteins from the trypsin solution, then resuspended in 1 ml PBS. 20% of each sample was transferred to a new 1.5 ml DNA LoBind tube tube for downstream isolation of RNA and protein. Cells transfected with Luciferase siRNA or Negative Control No. 2 siRNA were used as negative controls. RNA was extracted from 20% of the primary hepatocytes in each transfected 6-well plate well using a Quick-RNA Microprep Kit. Library preparation was performed using the NEBNext Ultra Express RNA Library Prep Kit and NEBNext Poly(A) mRNA Magnetic Isolation Module.
ENA first public
2026-06-11
INSDC center name
German Cancer Research Center
INSDC status
public
Submitter Id
E-MTAB-17079:Sample 10
broker name
ArrayExpress
cell line
primary hepatocytes
cell type
hepatocytes
collection date
not collected
common name
western European house mouse
developmental stage
adult
disease
normal
genotype
wild type genotype
geographic location (country and/or sea)
not collected
isolate
not applicable
organism part
liver
replicate
2
sample name
E-MTAB-17079:Sample 10
scientific_name
Mus musculus domesticus