2024-04-01
ENA first public
ERC000011
ENA-CHECKLIST
Norwegian University of Science and Technology (NTNU) Department of Circulation and Medical Imaging
INSDC center name
public
INSDC status
ERS18433042
SRA accession
E-MTAB-13912:MEA_CONTROL_RNA_1
Submitter Id
ArrayExpress
broker name
Ncyte® vCardiomyocytes
cell_line
induced pluripotent stem cell
cell_type
not collected
collection date
human
common name
Protocols: Cells, in 96-well plate, were washed gently with PBS (1X) before QIAzol lysis reagent (100 uL) (Qiagen, #79306, Hilden, Germany) was added to the cells and the lysate was collected and pooled in to three samples for the control group and three samples for the EXO-preconditioned group   Ncyte™ Cardioplate™ Maestro™ MEA 96 with pre-seeded IPSC-cardiomyocytes (Ncardia, # Nc-K-NCP-M96, Leiden, NL) was maintained in in Cardiomyocyte Culture Media (Nc-M-CMCM-250, Ncardia, Leiden, NL) at 21% O2, 5% CO2 in a humidified atmosphere at 37C, until stable electrophysiological activity was observed using the Multi-electrode array (MEA) systems. exosome-precondition group (MEA_EXO_RNA) was administred exosomes 24 hpurs before 16 hours of hypoxic stimulus. MEA_CONTROL_RNA was not exposed to exosomes (only medium change) 24 hours before 16 hours of hypoxic stimulus.  RNA was isolated using the miRNeasy Micro Kit (Qiagen, #217084, Hilden, Germany) following the manufacturer’s instructions. RNA sample quality control was performed using the RNA 6000 Pico Kit (Agilent, #5067-1513, Santa Clara, CA, USA) with a 2100 Bioanalyzer System (Agilent, #G2939BA, Santa Clara, CA, USA) following the manufacturer’s instructions. Then, a NEXTFLEX® Small RNA-Seq Kit v4 with UDIs (PerkinElmer, #NOVA-5132-44 and #NOVA-5132-32, Waltham, Massachusetts, USA) was used to generate the sample library, and a 4200 TapeStation system (Agilent, Santa Clara, CA, USA) was used for quality control of the sample library. Based on the TapeStation data, sample concentrations were adjusted and then quantified using a Qubit Q32853 with the Qubit™ dsDNA BR Assay Kit (Invitrogen, #Q32850, Waltham, Massachusetts, USA).
description
not collected
geographic location (country and/or sea)
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
cardiac ventricle
organism part
Homo sapiens
scientific_name
16h hypoxia
stimulus
MEA_CONTROL_RNA_1
title