2025-06-06
ENA first public
ERC000011
ENA-CHECKLIST
UCL
INSDC center name
public
INSDC status
ERS21187515
SRA accession
E-MTAB-14537:BDNF_1F_6hr
Submitter Id
ArrayExpress
broker name
WTC-11
cell_line
iPSC Lower Motor Neuron
cell_type
not collected
collection date
human
common name
Protocols: The human induced pluripotent stem cells used are the WTC11 line, stably expressing a doxycycline-inducible hNIL construct and CRISPR/cas9. Briefly, iPSCs were seeded on tissue culture dishes coated with 120-180 μg/ml Geltrex (Thermo Fisher). Cells were maintained in iPSC medium containing Essential 8 Flex Medium (E8F, Thermo Fisher), E8 Flex supplement (Thermo Fisher), and 100 U/ml Pen/Strep and maintained in a 37 ºC, 5% CO2 incubator. Medium change was performed every 1-2 days. Passage of cells were performed with Stem Pro Accutase (Thermo Fisher) for 1-2 min at 37 ºC. Accutase was removed by centrifugation at 300 g for 5 min, and the cells were reseeded in iPS cell media supplemented with 10 μM ROCK inhibitor (Y-27632, Selleckchem) to facilitate survival. ROCK inhibitor was removed after 24 h. Cells were washed with PBS and RNA was extracted using the Qiagen RNeasy isolation and purification kit. RNA concentration was measured, and after equilibrating the amount of RNA in each sample, iodoacetamide (10 mM), NaPOH4 (pH 8, 50 mM), and DMSO (50% v/v) were added to each sample and incubated at 50 C for 15 min to facilitate the thiol-alkylation of 4sU.  Sequencing libraries were prepared using the KAPA HyperPrep Kit with RiboErase kit (Roche).
description
adult
developmental stage
normal
disease
wild type genotype
genotype
not collected
geographic location (country and/or sea)
DIV 7
growth condition
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
blood
organism part
Homo sapiens
scientific_name
BDNF
stimulus
BDNF_1F_6hr
title