發(fā)布時間：2018-04-29 18:09

  本文選題：聲帶 + 創(chuàng)傷和損傷��； 參考：《復旦大學》2012年碩士論文

【摘要】：目的研究人羊膜上皮細胞(human amniotic epithelial cells, hAECs)移植入兔損傷聲帶組織內(nèi)的生長分布特點,探索hAECs促進損傷聲帶修復再生的潛能。方法分離和培養(yǎng)人羊膜上皮細胞,利用慢病毒載體將增強型綠色熒光蛋白(enhanced green fluorescent protein, EGFP)基因轉染至hAECs內(nèi)。建立深及聲韌帶的兔聲帶損傷模型,設hAECs移植組即實驗組(13側聲帶)、損傷對照組(13側聲帶)及正常對照組(4側聲帶)。熒光顯微鏡下連續(xù)觀察hAECs在聲帶內(nèi)的存活、分布情況,免疫熒光染色觀察hAECs在聲帶固有層內(nèi)的分化情況。應用HE染色和免疫組化染色分析膠原纖維、纖維連接蛋白等主要細胞外基質(zhì)在損傷后3月時的含量及分布。結果hAECs原代培養(yǎng)6d后呈鋪路石樣生長,移植入兔損傷聲帶組織后可在固有層內(nèi)持續(xù)存活2個月,細胞呈縱向排列,有趨向性。聲帶損傷2月時,免疫熒光顯示hAECs移植組肌細胞標志蛋白Desmin熒光陽性,提示hAECs向肌細胞分化。光鏡觀察下見hAECs移植后3個月,兔聲帶內(nèi)膠原纖維密度和排布較損傷對照組改善,但未及正常。免疫組化染色示hAECs移植組纖維連接蛋白含量和分布介于損傷對照組和正常對照組之間。結論hAECs可在異種動物聲帶損傷組織內(nèi)持續(xù)存活、生長,并有向聲帶組織分化的潛能,可促進聲帶損傷后的修復再生。
[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.
【學位授予單位】：復旦大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R767.4

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相關期刊論文 前2條

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本文編號：1821013