重组腺病毒Ad-GFP转染新生小鼠离体耳蜗基底膜的实验分析

目的观察重组腺病毒Ad-GFP转染新生小鼠离体耳蜗基底膜培养组织的情况。方法取新生1～5 d小鼠的耳蜗基底膜,离体培养1 d后,加入重组腺病毒Ad-GFP继续培养1 d,在荧光显微镜及Confocal显微镜下观察病毒对离体耳蜗基底膜的转染情况。结果耳蜗基底膜离体培养1 d后,在显微镜下观察,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。离体耳蜗基底膜培养组织加入重组腺病毒Ad-GFP继续培养1 d后,可见重组腺病毒Ad-GFP能高效转染新生小鼠离体耳蜗基底膜及其外周长出的新生上皮细胞和成纤维细胞;在新生小鼠离体耳蜗基底膜上,不仅大上皮嵴细胞区域、小上皮嵴细胞区域的细胞能被高效转染,毛细胞也能被高效转染。结论重组腺病毒Ad-GFP能高效转染新生小鼠离体耳蜗基底膜培养组织。

Recombinant adenovirus transfects organ of Corti from neonatal mouse in vitro

Objective To detect the transfection of organ of Corti from neonatal mouse with Ad-GFP in vitro. Methods One day after tissue cultivation, the organ of Corti from neonatal mouse was transfeeted with Ad-GFP in vitro. One day after transfection, the transfection efficiency was analyzed with Confocal and fluorescence microscopes. Results One day after the tissue cultivation, the organ of Corti grew wel 1 and new epithelial cells and fibroblasts were found around the tissue. One day after the transfection, the hair cells, the cells in lesser and greater epithelial ridges, the new epithelial cells and fibroblasts around the tissue were effectively transfeeted by Ad-GFP. Conclusions Ad-GFP can transfect organ of Corti from neonatal mouse effectively in vitro.