From Otic Induction to Hair Cell Production: Pax2EGFP Cell Line Illuminates Key Stages of Development in Mouse Inner Ear Organoid Model
| Autor: | Thomas Schrepfer, Stacy A Schaefer, Gabriel Corfas, Robert Keith Duncan, Guoqiang Wan, Atsuko Y. Higashi, Benjamin R. Loomis, Gregory R. Dressler |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
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 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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