Generation of inner ear sensory epithelia from pluripotent stem cells in 3D culture
Open accessAbstract
The inner ear contains sensory epithelia that detect head movements, gravity and sound. It is unclear how to develop these sensory epithelia from pluripotent stem cells, a process that will be critical for modelling inner ear disorders or developing cell-based therapies for profound hearing loss and balance disorders. So far, attempts to derive inner ear mechanosensitive hair cells and sensory neurons have resulted in inefficient or incomplete phenotypic conversion of stem cells into inner-ear-like cells. A key insight lacking from these previous studies is the importance of the non-neural and preplacodal ectoderm, two critical precursors during inner ear development. Here we report the stepwise differentiation of inner ear sensory epithelia from mouse embryonic stem cells (ESCs) in three-dimensional culture. We show that by recapitulating in vivo development with precise temporal control of signalling pathways, ESC aggregates transform sequentially into non-neural, preplacodal and otic-placode-like epithelia. Notably, in a self-organized process that mimics normal development, vesicles containing prosensory cells emerge from the presumptive otic placodes and give rise to hair cells bearing stereocilia bundles and a kinocilium. Moreover, these stem-cell-derived hair cells exhibit functional properties of native mechanosensitive hair cells and form specialized synapses with sensory neurons that have also arisen from ESCs in the culture. Finally, we demonstrate how these vesicles are structurally and biochemically comparable to developing vestibular end organs. Our data thus establish a new in vitro model of inner ear differentiation that can be used to gain deeper insight into inner ear development and disorder.
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TGFβ inhibition promotes ectoderm induction from pluripotent stem cells.
“TGFβ inhibition promotes ectoderm induction from pluripotent stem cells, whereas BMP activation promotes surface ectoderm (alternatively, non-neural ectoderm) induction and suppresses neuroectoderm induction9–12.”
From Hair-bearing human skin generated entirely from pluripotent stem cells (2020) · cites this paper as [10]
The paper’s concluding passage (Abstract; page unavailable) says TGFβ inhibition acts with BMP activation in the induction of non-neural ectoderm in vitro. It does not establish that TGFβ inhibition alone promotes ectoderm induction from pluripotent stem cells, as the cited clause could imply. (LLM, conf=0.84)
Experimental — accuracy not yet measuredSB/BMP treated aggregates, without subsequent FGF and LDN treatment, produced epidermal keratinocyte cysts largely devoid of surrounding mesenchymal tissue.
“We knew from our previous work that SB/BMP treated aggregates, without subsequent FGF and LDN treatment, produced epidermal keratinocyte cysts largely devoid of surrounding mesenchymal tissue (Koehler et al., 2013).”
From Hair Follicle Development in Mouse Pluripotent Stem Cell-Derived Skin Organoids (2018) · cites this paper as Koehler et al., 2013
The passage on BMP/SB-treated aggregates (section labeled Abstract; no page provided) says their outer epithelium develops into Krt5/p63+ epidermal epithelium, supporting the epidermal-fate portion of the claim. It does not say that these aggregates form keratinocyte cysts or are largely devoid of surrounding mesenchymal tissue. (LLM, conf=0.83)
Experimental — accuracy not yet measuredIn the 3D mESC culture, a heterogeneous population of mesenchymal and neural cells is also generated.
“In the process of culture, a heterogeneous population of mesenchymal and neural cells is also generated (Koehler and Hashino, 2014; Koehler et al., 2013).”
From Hair Follicle Development in Mouse Pluripotent Stem Cell-Derived Skin Organoids (2018) · cites this paper as Koehler et al., 2013
The cellular-composition passage (indexed under Abstract; page unavailable) describes day-5 mouse ESC aggregates as containing a mixture of mesendodermal and neuroectodermal tissues, supporting the claim that the culture generates heterogeneous cells including neural-lineage cells. It does not identify mesenchymal cells; mesendodermal and mesenchymal are not interchangeable. (LLM, conf=0.87)
Experimental — accuracy not yet measuredInduced surface epithelia generate p63+ KRT5+ basal keratinocyte-like cells that self-organize into a cyst, referred to as a skin organoid.
“In addition, we also briefly described how the induced surface epithelia generate p63+ Keratin 5 (KRT5)+ basal keratinocyte-like cells that self-organize into a cyst (Koehler et al., 2013), referred to hereafter as a skin organoid.”
From Hair Follicle Development in Mouse Pluripotent Stem Cell-Derived Skin Organoids (2018) · cites this paper as Koehler et al., 2013
The passage on BMP/SB-treated aggregates (section labeled Abstract; page unavailable) says the outer epithelium develops into Krt5/p63+ epithelium resembling epidermis. A second passage describes a basal p63+ layer, but the copy does not establish that these cells self-organize into a cyst. The term “skin organoid” is introduced by the citing authors, not attributed to Koehler et al. (LLM, conf=0.86)
Experimental — accuracy not yet measuredTo successfully generate skin, co-development of epidermis and dermis is required, and precise alterations of major signaling pathways (TGF-β, BMP4, and bFGF) are critical.
“To successfully generate skin, co-development of epidermis and dermis is required, and precise alterations of major signaling pathways (TGF-β, BMP4, and bFGF) are critical”
From Generation and characterization of hair-bearing skin organoids from human pluripotent stem cells (2022) · cites this paper as [30]
The paper’s Abstract describes timed changes to TGFβ, BMP, and FGF signaling to generate inner-ear precursors, and reports epidermis-like epithelium in some aggregates. It does not say that these changes are critical for generating skin or that epidermis and dermis must co-develop; the cited result concerns inner-ear development. (LLM, conf=0.86)
Experimental — accuracy not yet measured
BMP activation promotes surface ectoderm induction and suppresses neuroectoderm induction.
“TGFβ inhibition promotes ectoderm induction from pluripotent stem cells, whereas BMP activation promotes surface ectoderm (alternatively, non-neural ectoderm) induction and suppresses neuroectoderm induction9–12.”
From Hair-bearing human skin generated entirely from pluripotent stem cells (2020) · cites this paper as [10]
A 3D mouse embryonic stem cell culture can produce cranial surface epithelia (non-neural ectoderm), a precursor tissue of the skin epidermis.
“Recently, we developed a three-dimensional (3D) mouse embryonic stem cell (mESC) culture capable of producing cranial surface epithelia (also known as non-neural ectoderm), a precursor tissue of the skin epidermis.”
From Hair Follicle Development in Mouse Pluripotent Stem Cell-Derived Skin Organoids (2018) · cites this paper as Koehler et al., 2013
Culture systems containing epidermal and dermal cells have been used to model human skin outside of the body for research or clinical uses for over 40 years.
“Culture systems containing epidermal and dermal cells have been used to model human skin outside of the body for research or clinical uses for over 40 years7,8,10; however, growing functional skin appendages in these systems has been persistently difficult.”
From Hair-bearing human skin generated entirely from pluripotent stem cells (2020) · cites this paper as [10]
The paper’s Abstract describes a 3D culture model of inner-ear development and notes an epithelium that mimics embryonic epidermis. Our copy does not address culture systems combining epidermal and dermal cells to model human skin, their research or clinical uses, or a 40-year history (searched for skin culture and its history). (LLM, conf=0.77)
Experimental — accuracy not yet measuredGrowing functional skin appendages in culture systems containing epidermal and dermal cells has been persistently difficult.
“Culture systems containing epidermal and dermal cells have been used to model human skin outside of the body for research or clinical uses for over 40 years7,8,10; however, growing functional skin appendages in these systems has been persistently difficult.”
From Hair-bearing human skin generated entirely from pluripotent stem cells (2020) · cites this paper as [10]
The paper’s Abstract describes generating inner-ear sensory epithelia and mechanosensitive hair cells from mouse embryonic stem cells. Searches for skin appendages and epidermal–dermal culture systems found no passage addressing whether functional skin appendages are difficult to grow in those systems. The paper’s inner-ear hair cells are not skin appendages. (LLM, conf=0.77)
Experimental — accuracy not yet measured
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- Authors
- Koehler, K. R., Mikosz, A. M., Molosh, A. I., Patel, D., & Hashino, E.
- Affiliations
- Indiana University Indianapolis (US)
- Corresponding author
- Koehler, Karl R.
- Hashino, Eri
- Venue
- Nature
- Published
- 2013-07-10
- Volume
- 500
- Issue
- 7461
- Pages
- 217-221
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- PMC
- PMC3739998
- OpenAlex
- W1967800133
- Cited by
- 410
- References
- 3
- Citations by year
- 2026: 16 · 2025: 28 · 2024: 20 · 2023: 24 · 2022: 43 · 2021: 38 · 2020: 31 · 2019: 27 · 2018: 31 · 2017: 35 · 2016: 35 · 2015: 37 · 2014: 38 · 2013: 6 · 2012: 1
- FWCI
- 22.30
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Nature (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- IUScholarWorks (Indiana University) (submittedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- Funders
- Indiana Clinical and Translational Sciences Institute
- National Institutes of Health (R01MH52619)
- School of Medicine, Indiana University
- National Institute of Mental Health (R01-MH52619, R01MH052619)
- National Institute on Deafness and Other Communication Disorders (R21DC012617, RC1DC010706)
- National Institute of General Medical Sciences (R01GM086544)
- National Center for Research Resources (TL1RR025759)
- Topics
- Life Sciences › Neuroscience › Sensory Systems › Hearing, Cochlea, Tinnitus, Genetics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
- Keywords
- mouse embryonic stem cells, 3D cell culture, otic placode, stereocilia, hair cell synapses, vestibular organs, inner ear development, hearing loss, balance disorders, disease modeling, cell therapy, in vitro differentiation model, kinocilium
- MeSH terms
- Myosin VIIa, Animals, Cell Differentiation, Hair Cells, Auditory, Inner, Humans, Ear, Inner, Myosins, Recombinant Proteins, Gene Expression Regulation, Developmental, Cell Culture Techniques, Pluripotent Stem Cells, Mice, Electrical Synapses, Neurogenesis
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