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From Otic Induction to Hair Cell Production: Pax2EGFP Cell Line Illuminates Key Stages of Development in Mouse Inner Ear Organoid Model
- Publication Year :
- 2018
- Publisher :
- Mary Ann Liebert, Inc., 2018.
-
Abstract
- Producing hair cells of the inner ear is the major goal of ongoing research that combines advances in developmental and stem cell biology. The recent advent of an inner ear organoid protocol-resulting in three-dimensional stem cell-derived tissues resembling vestibular sensory epithelia-has sparked interest in applications such as regeneration, drug discovery, and disease modeling. In this study, we adapted this protocol for a novel mouse embryonic stem cell line with a fluorescent reporter for Pax2 expression. We used Pax2EGFP/+ organoid formation to model otic induction, the pivotal developmental event when preplacodal tissue adopts otic fate. We found upregulation of Pax2 and activation of ERK downstream of fibroblast growth factor signaling in organoid formation as in embryonic inner ear development. Pax2 expression was evident from the EGFP reporter beginning at the vesicle formation stage and persisting through generation of the sensory epithelium. The native ventralizing signal sonic hedgehog was largely absent from the cell aggregates as otic vesicles began to form, confirming the dorsal vestibular organoid fate. Nonetheless, cochlear- or vestibular-like neurons appeared to delaminate from the derived otic vesicles and formed synaptic contacts with hair cells in the organoids. Cell lines with transcriptional reporters such as Pax2EGFP/+ facilitate direct evaluation of morphological changes during organoid production, a major asset when establishing and validating the culture protocol.
- Subjects :
- 0301 basic medicine
Organogenesis
Green Fluorescent Proteins
Mice, Transgenic
Biology
Fibroblast growth factor
Cell Line
03 medical and health sciences
Mice
Original Research Reports
Hair Cells, Auditory
medicine
Organoid
otorhinolaryngologic diseases
Animals
Inner ear
Sonic hedgehog
Regeneration (biology)
PAX2 Transcription Factor
Gene Expression Regulation, Developmental
Cell Differentiation
Mouse Embryonic Stem Cells
Cell Biology
Hematology
Embryonic stem cell
Cell biology
Mice, Inbred C57BL
Organoids
030104 developmental biology
medicine.anatomical_structure
Microscopy, Fluorescence
Cell culture
Ear, Inner
biology.protein
Hair cell
sense organs
Developmental Biology
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Accession number :
- edsair.doi.dedup.....f3f9ebd6ff40c1268eff32f484a42cbd