人羊膜上皮细胞移植用于兔损伤声带组织修复的初步研究

发布时间：2018-04-29 18:09

  本文选题：声带 + 创伤和损伤　； 参考：《复旦大学》2012年硕士论文

【摘要】：目的研究人羊膜上皮细胞(human amniotic epithelial cells, hAECs)移植入兔损伤声带组织内的生长分布特点,探索hAECs促进损伤声带修复再生的潜能。方法分离和培养人羊膜上皮细胞,利用慢病毒载体将增强型绿色荧光蛋白(enhanced green fluorescent protein, EGFP)基因转染至hAECs内。建立深及声韧带的兔声带损伤模型,设hAECs移植组即实验组(13侧声带)、损伤对照组(13侧声带)及正常对照组(4侧声带)。荧光显微镜下连续观察hAECs在声带内的存活、分布情况,免疫荧光染色观察hAECs在声带固有层内的分化情况。应用HE染色和免疫组化染色分析胶原纤维、纤维连接蛋白等主要细胞外基质在损伤后3月时的含量及分布。结果hAECs原代培养6d后呈铺路石样生长,移植入兔损伤声带组织后可在固有层内持续存活2个月,细胞呈纵向排列,有趋向性。声带损伤2月时,免疫荧光显示hAECs移植组肌细胞标志蛋白Desmin荧光阳性,提示hAECs向肌细胞分化。光镜观察下见hAECs移植后3个月,兔声带内胶原纤维密度和排布较损伤对照组改善,但未及正常。免疫组化染色示hAECs移植组纤维连接蛋白含量和分布介于损伤对照组和正常对照组之间。结论hAECs可在异种动物声带损伤组织内持续存活、生长,并有向声带组织分化的潜能,可促进声带损伤后的修复再生。
[Abstract]:Objective to study the growth and distribution of human amniotic epithelial cells (hAECs) transplanted into injured vocal cord tissues of rabbits, and to explore the potential of hAECs to promote the repair and regeneration of injured vocal cords. Methods Human amniotic epithelial cells were isolated and cultured. The enhanced green fluorescent protein enhanced green fluorescent protein (EGFPFP) gene was transfected into hAECs by lentivirus vector. The rabbit vocal cord injury model with deep and acoustical ligament was established. The experimental group was divided into experimental group (13 sides) and the control group (13 sides) and the normal control group (4 sides of vocal cord). The survival and distribution of hAECs in vocal cord were observed continuously under fluorescence microscope, and the differentiation of hAECs in lamina propria of vocal cord was observed by immunofluorescence staining. The content and distribution of collagen fibers, fibronectin and other major extracellular matrix were analyzed by HE staining and immunohistochemical staining at 3 months after injury. Results after 6 days of primary culture, hAECs grew like paving stone. After transplantation into rabbit vocal cord tissue, the cells could survive in the lamina propria for 2 months, and the cells were in a longitudinal arrangement and tended to be in the lamina propria. After 2 months of vocal cord injury, immunofluorescence showed that Desmin was positive in hAECs transplantation group, suggesting that hAECs differentiated into myocytes. Three months after hAECs transplantation, the density and arrangement of collagen fibers in the vocal cords of rabbits were better than those in the injured control group, but they were not normal. Immunohistochemical staining showed that the content and distribution of fibronectin in hAECs transplantation group were between the injury control group and the normal control group. Conclusion hAECs can survive, grow and differentiate into vocal cord tissue of xenogeneic animals. It can promote the repair and regeneration after vocal cord injury.
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R767.4

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