20pcw-84
Homo sapiens
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.
ENA first public
2024-11-04
INSDC center name
1. UCL Great Ormond Street Institute of Child Health, University College London, and NIHR Great Ormond Street Hospital Biomedical Research Centre, London, UK.
INSDC status
public
Submitter Id
E-MTAB-14063:20pcw-84
age
20
broker name
ArrayExpress
cell_type
multiple
collection date
not collected
common name
human
developmental stage
20 post conception weeks
disease
normal
geographic location (country and/or sea)
not collected
individual
84
isolate
not applicable
organism part
cochlea
sample name
E-MTAB-14063:20pcw-84
scientific_name
Homo sapiens
sex
unknown