Modulation of Wnt Signaling Enhances Inner Ear Organoid Development in 3D Culture.
Stem cell-derived inner ear sensory epithelia are a promising source of tissues for treating patients with hearing loss and dizziness. We recently demonstrated how to generate inner ear sensory epithelia, designated as inner ear organoids, from mouse embryonic stem cells (ESCs) in a self-organizing 3D culture. Here we improve the efficiency of this culture system by elucidating how Wnt signaling activity can drive the induction of otic tissue. We found that a carefully timed treatment with the potent Wnt agonist CHIR99021 promotes induction of otic vesicles-a process that was previously self-organized by unknown mechanisms. The resulting otic-like vesicles have a larger lumen size and contain a greater number of Pax8/Pax2-positive otic progenitor cells than organoids derived without the Wnt agonist. Additionally, these otic-like vesicles give rise to large inner ear organoids with hair cells whose morphological, biochemical and functional properties are indistinguishable from those of vestibular hair cells in the postnatal mouse inner ear. We conclude that Wnt signaling plays a similar role during inner ear organoid formation as it does during inner ear development in the embryo.
Department of Otolaryngology-Head and Neck Surgery, Indiana University School of Medicine, Indianapolis, IN, 46202, United States of America.|Stark Neurosciences Research Institute, Indiana University School of Medicine, Indianapolis, IN, 46202, United States of America.
Rachel E
RE
DeJonge
Department of Otolaryngology, F.M. Kirby Neurobiology Center Boston Children's Hospital, and Harvard Medical School, Boston, MA, 02115, United States of America.
Xiao-Ping
XP
Liu
Department of Otolaryngology-Head and Neck Surgery, Indiana University School of Medicine, Indianapolis, IN, 46202, United States of America.
Christopher R
CR
Deig
Department of Otolaryngology-Head and Neck Surgery, Stanford University, Palo Alto, CA, 94305, United States of America.
Stefan
S
Heller
Department of Otolaryngology-Head and Neck Surgery, Indiana University School of Medicine, Indianapolis, IN, 46202, United States of America.|Stark Neurosciences Research Institute, Indiana University School of Medicine, Indianapolis, IN, 46202, United States of America.
Karl R
KR
Koehler
Department of Otolaryngology-Head and Neck Surgery, Indiana University School of Medicine, Indianapolis, IN, 46202, United States of America.|Stark Neurosciences Research Institute, Indiana University School of Medicine, Indianapolis, IN, 46202, United States of America.
Eri
E
Hashino
D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism; D002449:Cell Aggregation / Q000187:drug effects; D007758:Ear, Inner / Q000187:drug effects / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D040542:Mechanotransduction, Cellular / Q000187:drug effects; D051379:Mice; D000081412:Myosin VIIa; D009218:Myosins / Q000378:metabolism; D009940:Organoids / Q000187:drug effects / Q000378:metabolism*; D039904:Pluripotent Stem Cells / Q000187:drug effects / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D046509:Tissue Culture Techniques / Q000379:methods*; D060449:Wnt Signaling Pathway / Q000187:drug effects
D051792:Basic Helix-Loop-Helix Proteins; D049452:Green Fluorescent Proteins; D000081412:Myosin VIIa; D009218:Myosins; D011725:Pyridines; D011743:Pyrimidines; D055748:SOXB1 Transcription Factors; C500943:Atoh1 protein, mouse; C473711:Chir 99021; C000631455:MYO7A protein, human; C000631456:Myo7a protein, mouse