PMID- 29311837
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20200930
IS  - 1662-5102 (Print)
IS  - 1662-5102 (Electronic)
IS  - 1662-5102 (Linking)
VI  - 11
DP  - 2017
TI  - Generation of Otic Sensory Neurons from Mouse Embryonic Stem Cells in 3D Culture.
PG  - 409
LID - 10.3389/fncel.2017.00409 [doi]
LID - 409
AB  - The peripheral hearing process taking place in the cochlea mainly depends on two 
      distinct sensory cell types: the mechanosensitive hair cells and the spiral 
      ganglion neurons (SGNs). The first respond to the mechanical stimulation exerted 
      by sound pressure waves on their hair bundles by releasing neurotransmitters and 
      thereby activating the latter. Loss of these sensorineural cells is associated 
      with permanent hearing loss. Stem cell-based approaches aiming at cell 
      replacement or in vitro drug testing to identify potential ototoxic, 
      otoprotective, or regenerative compounds have lately gained attention as putative 
      therapeutic strategies for hearing loss. Nevertheless, they rely on efficient and 
      reliable protocols for the in vitro generation of cochlear sensory cells for 
      their implementation. To this end, we have developed a differentiation protocol 
      based on organoid culture systems, which mimics the most important steps of in 
      vivo otic development, robustly guiding mouse embryonic stem cells (mESCs) toward 
      otic sensory neurons (OSNs). The stepwise differentiation of mESCs toward 
      ectoderm was initiated using a quick aggregation method in presence of Matrigel 
      in serum-free conditions. Non-neural ectoderm was induced via activation of bone 
      morphogenetic protein (BMP) signaling and concomitant inhibition of transforming 
      growth factor beta (TGFbeta) signaling to prevent mesendoderm induction. Preplacodal 
      and otic placode ectoderm was further induced by inhibition of BMP signaling and 
      addition of fibroblast growth factor 2 (FGF2). Delamination and differentiation 
      of SGNs was initiated by plating of the organoids on a 2D Matrigel-coated 
      substrate. Supplementation with brain-derived neurotrophic factor (BDNF) and 
      neurotrophin-3 (NT-3) was used for further maturation until 15 days of in vitro 
      differentiation. A large population of neurons with a clear bipolar morphology 
      and functional excitability was derived from these cultures. Immunostaining and 
      gene expression analysis performed at different time points confirmed the 
      transition trough the otic lineage and final expression of the key OSN markers. 
      Moreover, the stem cell-derived OSNs exhibited functional electrophysiological 
      properties of native SGNs. Our established in vitro model of OSNs development can 
      be used for basic developmental studies, for drug screening or for the 
      exploration of their regenerative potential.
FAU - Perny, Michael
AU  - Perny M
AD  - Neuroinfection Laboratory, Institute for Infectious Diseases, University of Bern, 
      Bern, Switzerland.
AD  - Laboratory of Inner Ear Research, Department for BioMedical Research, University 
      of Bern, Bern, Switzerland.
AD  - Department of Otorhinolaryngology, Head and Neck Surgery, Inselspital, University 
      of Bern, Bern, Switzerland.
AD  - Cluster for Regenerative Neuroscience, Department of Biomedical Research, 
      University of Bern, Bern, Switzerland.
FAU - Ting, Ching-Chia
AU  - Ting CC
AD  - Laboratory of Inner Ear Research, Department for BioMedical Research, University 
      of Bern, Bern, Switzerland.
AD  - Department of Otorhinolaryngology, Head and Neck Surgery, Inselspital, University 
      of Bern, Bern, Switzerland.
AD  - Cluster for Regenerative Neuroscience, Department of Biomedical Research, 
      University of Bern, Bern, Switzerland.
FAU - Kleinlogel, Sonja
AU  - Kleinlogel S
AD  - Institute for Physiology, University of Bern, Bern, Switzerland.
FAU - Senn, Pascal
AU  - Senn P
AD  - Laboratory of Inner Ear Research, Department for BioMedical Research, University 
      of Bern, Bern, Switzerland.
AD  - Department of Otorhinolaryngology, Head and Neck Surgery, Inselspital, University 
      of Bern, Bern, Switzerland.
AD  - Cluster for Regenerative Neuroscience, Department of Biomedical Research, 
      University of Bern, Bern, Switzerland.
AD  - Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital 
      Geneva (HUG), Geneva, Switzerland.
FAU - Roccio, Marta
AU  - Roccio M
AD  - Laboratory of Inner Ear Research, Department for BioMedical Research, University 
      of Bern, Bern, Switzerland.
AD  - Department of Otorhinolaryngology, Head and Neck Surgery, Inselspital, University 
      of Bern, Bern, Switzerland.
AD  - Cluster for Regenerative Neuroscience, Department of Biomedical Research, 
      University of Bern, Bern, Switzerland.
LA  - eng
PT  - Journal Article
DEP - 20171219
PL  - Switzerland
TA  - Front Cell Neurosci
JT  - Frontiers in cellular neuroscience
JID - 101477935
PMC - PMC5742223
OTO - NOTNLM
OT  - 3D culture
OT  - in vitro mESC differentiation
OT  - organoids
OT  - otic development
OT  - spiral ganglion neurons
EDAT- 2018/01/10 06:00
MHDA- 2018/01/10 06:01
PMCR- 2017/01/01
CRDT- 2018/01/10 06:00
PHST- 2017/09/22 00:00 [received]
PHST- 2017/12/05 00:00 [accepted]
PHST- 2018/01/10 06:00 [entrez]
PHST- 2018/01/10 06:00 [pubmed]
PHST- 2018/01/10 06:01 [medline]
PHST- 2017/01/01 00:00 [pmc-release]
AID - 10.3389/fncel.2017.00409 [doi]
PST - epublish
SO  - Front Cell Neurosci. 2017 Dec 19;11:409. doi: 10.3389/fncel.2017.00409. 
      eCollection 2017.