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诱导多能干细胞体外诱导分化为耳蜗毛细胞样细胞和螺旋神经节神经元样细胞的研究

[Study on the induced differentiation of induced pluripotent stem cells into cochlear hair cell-like cells and spiral ganglion neuron-like cells in vitro].

作者信息

Guan Lina, Chen Yanhong, Zhu Hengtao, Chen Jing, Jiang Hongqun

机构信息

Department of Otorhinolaryngology Head and Neck Surgery, First Affiliated Hospital of Nanchang University, Nanchang 330006, China.

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出版信息

Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2014 Aug;49(8):680-6.

Abstract

OBJECTIVE

In this study, we investigated the potential of mouse induced pluripotent stem cells (iPSC) for use as a source of transplants for the restoration of auditory hair cells and spiral ganglion neurons.

METHODS

We co-cultured the mouse iPSC with the cells of the cochlear organ of Corti or the modiolus in vitro. The cochlear organ of Corti (which contains cochlear hair cells) and the modiolus (which contains auditory spiral ganglion neurons) were obtained from postnatal day 3 (P3) CD-1 ICR mice. After 18 days of coculture with the cells of newborn mouse cochleae. The expressions of hair cell markers (Myosin VIIa, Math1, Calretinin, Espin) and Spiral ganglion neuron markers [Nestin, Neurofilament-M, β-III Tubulin, Vesicular glutamate transporter 1(VGluT1)] were detected by immunocytochemical analysis.

RESULTS

Immunocytochemical analysis results indicated that the differentiated iPSC expressed auditory hair cell markers (MyosinVIIa,Math1, Calretinin, Espin ) and spiral ganglion markers (Nestin, Neurofilament-M,β-III Tubulin,VGluT1).

CONCLUSION

Mouse iPSC in virto cultured could successfully be induced to differentiate into hair cell-like cells and spiral ganglion-like cells with hair cell and spiral ganglion molecular markers.

摘要

目的

在本研究中,我们探究了小鼠诱导多能干细胞(iPSC)作为移植来源用于恢复听觉毛细胞和螺旋神经节神经元的潜力。

方法

我们将小鼠iPSC与来自柯蒂氏器或蜗轴的细胞在体外进行共培养。柯蒂氏器(包含耳蜗毛细胞)和蜗轴(包含听觉螺旋神经节神经元)取自出生后第3天(P3)的CD-1 ICR小鼠。与新生小鼠耳蜗细胞共培养18天后,通过免疫细胞化学分析检测毛细胞标志物(肌球蛋白VIIa、Math1、钙视网膜蛋白、肌动蛋白丝结合蛋白)和螺旋神经节神经元标志物[巢蛋白、神经丝蛋白-M、β-III微管蛋白、囊泡谷氨酸转运体1(VGluT1)]的表达。

结果

免疫细胞化学分析结果表明,分化的iPSC表达听觉毛细胞标志物(肌球蛋白VIIa、Math1、钙视网膜蛋白、肌动蛋白丝结合蛋白)和螺旋神经节标志物(巢蛋白、神经丝蛋白-M、β-III微管蛋白、VGluT1)。

结论

体外培养的小鼠iPSC能够成功诱导分化为具有毛细胞和螺旋神经节分子标志物的毛细胞样细胞和螺旋神经节样细胞。

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