Chen Yanni, Mu Wenbo, Wu Yongkang, Xu Jiake, Li Xiaofang, Hu Hui, Wang Siqi, Wang Dali, Hui Bin, Wang Lang, Dong Yi, Chen Wei
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.
College of Public Health, Shanghai University of Medicine & Health Sciences, Shanghai China.
J Tissue Eng. 2024 Jul 31;15:20417314241265198. doi: 10.1177/20417314241265198. eCollection 2024 Jan-Dec.
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 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.
螺旋神经节神经元（SGNs）损伤会导致耳聋。一种重要的治疗方法是利用干细胞来修复受损的听觉回路。然而，研究方法的实施不足给准确评估回路中衍生细胞的功能带来了挑战。在此，我们描述了一种将人类胚胎干细胞（hESCs）转化为耳神经元（ONs）的新方法，并使用光遗传学方法与共培养的细胞或大鼠耳蜗核（CN）的器官型切片来评估它们的功能连接性。胚胎干细胞衍生的耳神经元（eONs）在与耳蜗核神经元（CNNs）共培养时表现出SGN标记物表达并产生功能性突触连接。在脑片的耳蜗核中发现了突触素1和VGLUT表达，在eONs与CN脑片共培养期间，eONs在此处伸出突起。对eONs进行光刺激时，CNNs的动作电位尖峰和[此处原文缺失部分内容]同步增加。这些发现进一步证明eONs可能是治疗SGN耳聋的候选来源。
