characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2023-05-04T12:01:11Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2023-05-04T12:01:11Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA112194098
characteristics.characteristic: External Id
characteristics.text: The Francis Crick Institute UK Dementia Research Institute at King's College London
characteristics.characteristic: INSDC center name
characteristics.text: 2023-05-04T12:01:11Z
characteristics.characteristic: INSDC first public
characteristics.text: 2023-05-04T12:01:11Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS14305175
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-12402:WT_CT_1
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: NGN2 OPTi-OX hiPSC
characteristics.characteristic: cell_line
characteristics.text: cortical neuron
characteristics.characteristic: cell_type
characteristics.text: human
characteristics.characteristic: common name
characteristics.text: Protocols: The hIPSC cell line used has been published in the following paper: NGN2 OPTi-OX hiPSCs https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5390118/ . WT i-neurons in control (37 ºC throughout, collect at day 18), cooled (72 h at 32 ºC, day 15-18, collect at day 18) and rewarmed (72 h at 32 ºC, day 15-18, followed by 72 h at 37 ºC, day 18-21, collect at day 21) conditions were collected by detaching from tissue culture dishes. hiPSCs were enzymatically detached and dissociated into single cells using Accutase and plated into GelTrex (1:100 dilution)-coated culture plates in TeSR-E8 medium supplemented with 10 µM Rho-associated protein kinase (ROCK) inhibitor. After 24 hours (day 1), TeSR-E8 medium was changed to DMEM/F12 medium with GlutaMAX, supplemented with 1x N-2 supplement, 1x Non-Essential Amino Acids, 50 nM 2-Mercaptoethanol, 100 U/mL Penicillin-Streptomycin, and 1 µg/mL Doxycycline Hyclate (Dox) (iN-1 medium). After 24 hours (day 2), the medium was replaced with the same medium as the previous day. From day 3 to day 6, the culture was fed daily with Neurobasal medium supplemented with 1x B-27 supplement (minus vitamin A), 1x GlutaMAX, 50 nM 2-Mercaptoethanol, 100 U/mL Penicillin-Streptomycin, and 1 µg/mL Dox, 10 ng/mL Neurotrophin-3 (NT-3), and 10 ng/mL Brain-derived neurotrophic factor (BDNF) (iN-2 medium). After day 6, the medium was changed every other day. Total RNA was extracted with RNeasy Plus Mini Kit (Qiagen) following the manufacturer’s instructions. RNA concentrations of individual samples were measured using Qubit RNA Broad Range Assay Kit and their integrity was determined using Agilent TapeStation System. The library was prepared using Illumina TruSeq Stranded mRNA Library Prep following the manufacturer’s instructions.
characteristics.characteristic: description
characteristics.text: adult
characteristics.characteristic: developmental stage
characteristics.text: normal
characteristics.characteristic: disease
characteristics.text: wild type genotype
characteristics.characteristic: genotype
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: Homo sapiens
characteristics.characteristic: organism
characteristics.text: induced pluripotent stem cell
characteristics.characteristic: progenitor cell type
characteristics.text: Homo sapiens
characteristics.characteristic: scientific_name
characteristics.text: male
characteristics.characteristic: sex
characteristics.text: WT_CT_1
characteristics.characteristic: title