2024-11-04
ENA first public
ERC000011
ENA-CHECKLIST
1. UCL Great Ormond Street Institute of Child Health, University College London, and NIHR Great Ormond Street Hospital Biomedical Research Centre, London, UK.
INSDC center name
public
INSDC status
ERS19606130
SRA accession
E-MTAB-14063:20pcw-84
Submitter Id
20
week
age
ArrayExpress
broker name
multiple
cell_type
not collected
collection date
human
common name
Protocols: Human foetal cochlea samples were obtained from the Joint Medical Research Council UK/Wellcome Trust Human Developmental Biology Resource (http://www.hdbr.org/) with ethics approval.  On the day of collection, the cochlea was isolated from the temporal bone in PBS. The otic capsule was broken open and the membranous labyrinth and modiolus were isolated and transferred to a sterile tube containing 3 ml of enzyme mix (Miltenyi Neurosphere dissociation kit: 2880 µl Buffer X, 75 µl Enzyme P, 30 µl Buffer Y, 15 µl Enzyme A), incubated at 37°C for 60 minutes, rotating, followed by manual dissociation. The cell suspension was passed through a 40m cell strainer, diluted with an equal volume of 0.04% BSA-PBS. The cell suspension was centrifuged at 200 rcf (200 x g), 4°C for 5 minutes to pellet the cells. The supernatant was removed, and cells resuspended in 0.04% BSA-PBS and maintained on ice. Cells were quantified and cell viability assessed using Trypan blue stain on a haemocytometer. Samples with a viability of 65% or higher were used for scRNA sequencing. Single Cell RNA libraries were generated using the Chromium Single Cell 3’ Reagent Kit v3.1 (10X Genomics, CA, USA), with a target capture of ~10,000 cells per sample, following standard manufacturer’s specifications.
description
20 post conception weeks
developmental stage
normal
disease
not collected
geographic location (country and/or sea)
84
individual
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
cochlea
organism part
Homo sapiens
scientific_name
unknown
sex
20pcw-84
title