PDF(Pubmed)
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.
摘要：
螺旋神经节神经元（SGNs）损伤可引起耳聋。一种重要的治疗方法涉及利用干细胞来恢复受损的听觉电路。然而，研究方法的不充分实施对准确评估电路中衍生细胞的功能提出了挑战。这里,我们描述了一种将人胚胎干细胞（hESC）转化为耳神经元（ON）的新方法，并使用光遗传学方法与细胞或共培养大鼠耳蜗核（CN）的器官型切片评估其功能连接。与耳蜗核神经元（CNN）共培养时，胚胎干细胞衍生的耳神经元（eON）表现出SGN标记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的表达，在eON和CN脑切片共培养期间，eON预测了过程。CNN的动作电位尖峰和INa+/IK+随着对eON的光刺激而增加。这些发现提供了进一步的证据，证明eON可能是治疗SGN耳聋的候选来源。
