Optogenetically modified human embryonic stem cell-derived otic neurons establish functional synaptic connection with cochlear nuclei

Author:

Chen Yanni1234,Mu Wenbo1234,Wu Yongkang5,Xu Jiake6,Li Xiaofang7,Hu Hui8,Wang Siqi2,Wang Dali9,Hui Bin10,Wang Lang7,Dong Yi5,Chen Wei123411ORCID

Affiliation:

1. Institute of Translational Medicine, and Children’s Hospital Affiliated and Key Laboratory of Diagnosis and Treatment of Neonatal Diseases of Zhejiang Province, The Children’s Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, Hangzhou, China

2. College of Public Health, Shanghai University of Medicine & Health Sciences, Shanghai China

3. Institute of Wound Prevention and Treatment, Shanghai University of Medicine & Health Sciences, Shanghai, China

4. Institute of Translational Medicine, School of Medicine, Zhejiang University, Hangzhou, China

5. Key Laboratory of Adolescent Health Assessment and Exercise Intervention of Ministry of Education, School of Physical Education & Health Care, East China Normal University, Shanghai, China

6. Chongqing Key Laboratory of Translational Research for Cancer Metastasis and Individualized Treatment, Chongqing University Cancer Hospital, Chongqing, China

7. Department of Neurology of the First Affiliated Hospital, Interdisciplinary Institute of Neuroscience and Technology, Zhejiang University School of Medicine, Hangzhou, China

8. School of Laboratory Medicine and Bioengineering, Hangzhou Medical College, Hangzhou, China

9. Center for Clinical and Translational Medicine, Shanghai University of Medicine and Health Sciences, Shanghai, China

10. College of Pharmacy, Shanghai University of Medical & Health Sciences, Shanghai, China

11. Key Laboratory of Cellular Function and Pharmacology of Jilin Province, Yanbian University, Yanji, China

Abstract

Spiral ganglia neurons (SGNs) impairment can cause deafness. One important therapeutic approach involves utilizing stem cells to restore impaired auditory circuitry. Nevertheless, the inadequate implementation of research methodologies poses a challenge in accurately assessing the functionality of derived cells within the circuit. Here, we describe a novel method for converting human embryonic stem cells (hESCs) into otic neurons (ONs) and assess their functional connectivity using an optogenetic approach with cells or an organotypic slice of rat cochlear nucleus (CN) in coculture. Embryonic stem cell-derived otic neurons (eONs) exhibited SGN marker expression and generated functional synaptic connection when cocultured with cochlear nucleus neurons (CNNs). Synapsin 1 and VGLUT expression are found in the cochlear nucleus of brain slices, where eONs projected processes during the coculture of eONs and CN brain slices. Action potential spikes and INa+/IK+ of CNNs increased in tandem with light stimulations to eONs. These findings provide further evidence that eONs may be a candidate source to treat SGN-deafness.

Funder

natural science foundation of shanghai municipality

National Natural Science Foundation of China

Publisher

SAGE Publications

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