新生小鼠耳蜗基底膜体外培养的实验研究

In vitro cultivation of basilar membrane of cochlea from neonatal mouse

摘要:

目的建立可靠的新生小鼠离体耳蜗基底膜的培养方法，为内耳的研究提供新的条件和模型。方法在显微镜下完整分离出新生1～5 d小鼠的耳蜗基底膜组织，采用组织贴壁法在无血清培养基中进行培养，免疫荧光法检测新生小鼠离体耳蜗基底膜培养组织中毛细胞及螺旋神经元的生长状态。结果新生小鼠耳蜗基底膜离体培养24 h后，显微镜下观察可见基底膜贴壁生长良好，外周有新生上皮细胞和成纤维细胞长出；高倍显微镜下可见耳蜗内外毛细胞和支持细胞等结构；免疫荧光法检测可见毛细胞和螺旋神经元生长良好，并能保持较长一段时间。结论组织贴壁法无血清培养新生小鼠离体耳蜗基底膜是一种理想的耳科实验造模方法。

关键词:小鼠|耳蜗|柯替器|组织培养|毛细胞|螺旋神经元

Abstract:

Abstract:ObjectiveTo establish a reliable method for tissue culture of basilar membrane of cochlea from neonatal mouse.MethodsThe basilar membrane of cochlea was acquired from neonatal mouse with （the method of） microdissection. Then, the tissue was cultured in serumfree culture medium. Finally, the activities of the hair cells and the spiral ganglions were detected with immunofluorescence.ResultsTwentyfour hours after tissue cultivation, new epithelia and fibroblasts were found around the tissue. Inner and outer hair cells, supporting cells and spiral ganglions grew wel1 and could survive for a period of time.ConclusionAdherent cultivation of basilar membrane of cochlea in serumfree medium is an ideal method for otology research.

Key words:Mouse|Cochlea|Organ of Corti|Tissue cultivation|Hair cell|Spiral ganglion