organism: Homo sapiens
description: Protocols: Human cochlear organoids were generated from human pluripotent stem cell–based differentiation and served as the biological samples for this study. Organoids were maintained in CHIR99021-containing proliferative medium (EFI) and expanded for 10 days. Subsequently, organoids were subjected to adeno-associated virus (AAV)–mediated gene activation and cultured for an additional 7 days under differentiation conditions prior to sample collection.  At the designated collection time point, cochlear organoids were harvested and immediately transferred to ice-cold oxygenated artificial tissue preservation solution.  For scRNA-seq, harvested organoids were dissociated into single-cell suspensions by mincing and trypsin digestion at 37 °C for 20 min. After neutralization and filtration through a 40 μm strainer, cells were treated with RBC lysis buffer, washed, and resuspended in MEM supplemented with 1% BSA and RNase inhibitor. Cell concentration and viability were assessed using a LUNA-FL™ automated cell counter with LIVE/DEAD staining. For scRNA-seq, single-cell suspensions with viability exceeding 80% were used for scRNA-seq library preparation. Libraries were constructed according to the manufacturer's protocol using the Chromium Controller instrument and the Chromium Next GEM Single Cell 3′ Reagent Kits v3.1. Library preparation was carried out according to the manufacturer's protocol.
attributes.tag: ENA first public
attributes.value: 2026-02-11
attributes.tag: INSDC center name
attributes.value: Fudan
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-16612_2:AP_IRX2
attributes.tag: age
attributes.value: 7
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: mixed
attributes.tag: collection date
attributes.value: not collected
attributes.tag: common name
attributes.value: human
attributes.tag: developmental stage
attributes.value: none
attributes.tag: disease
attributes.value: normal
attributes.tag: geographic location (country and/or sea)
attributes.value: not collected
attributes.tag: isolate
attributes.value: not applicable
attributes.tag: organism part
attributes.value: cochlea organoids
attributes.tag: sample name
attributes.value: E-MTAB-16612_2:AP_IRX2
attributes.tag: scientific_name
attributes.value: Homo sapiens