Protocol for vibratome sectioning, immunofluorescence, and S-phase labeling of inner ear organoids.
Publication Year:
2025
PubMed ID:
40815565
Funding Grants:
Public Summary:
This study describes methods for analysis of inner ear sensory hair progenitor cells isolated from stem-cell-derived organoids. This enables the visualization of cell and tissue morphologies within developing organoids and assessment of how different manipulations affect cell proliferation, important for determining how mammalian inner ear sensory hair cells might be induced to repair after injury in order to restore hearing.
Scientific Abstract:
Inner ear organoids represent a potentially inexhaustible source of otic tissues, including sensory hair cells and supporting cells, for in vitro manipulation. Here, we present a protocol for labeling S-phase entry of cells in inner ear organoids using 5-ethynyl-2'-deoxyuridine (EdU), followed by fixation and vibratome sectioning. Nuclear EdU is then detected alongside protein markers of interest via immunofluorescence. This workflow enables the visualization of cell and tissue morphologies within developing organoids and assessment of how different manipulations affect cell proliferation. For complete details on the use and execution of this protocol, please refer to Matern et al.(1).
