单例先天性小耳残耳软骨细胞体外构建人耳廓软骨

发布时间：2018-05-20 04:40

  本文选题：先天性小耳畸形 + 残耳软骨细胞　； 参考：《基础医学与临床》2014年05期

【摘要】：目的探讨单例先天性小耳残耳软骨细胞体外构建正常人大小耳廓形态组织工程软骨的可行性。方法分离40例小耳畸形患者残耳软骨细胞统计细胞提取率;MTT法检测细胞增殖能力、计算扩增效率;免疫荧光和PCR检测不同代数细胞的软骨表型。分别应用3例患者各自的残耳软骨细胞培养至P3~P4代,藻酸盐凝胶包埋接种于正常人大小耳廓形态的聚羟基乙酸/聚乳酸支架,体外成软骨诱导动态培养10周后行组织学染色观察。结果耳软骨细胞提取率为(3.90±1.27)×106/g;在增殖培养基中细胞增殖能力明显提高,至P4代扩增(328.4±50.4)倍(P0.05);P3代细胞Ⅱ型胶原、蛋白聚糖表达显著减弱,至P4代消失,Ⅰ型胶原表达增强;体外培养10周,实验组形成了耳廓形态的类软骨组织,可见典型软骨陷窝,番红O、甲苯胺蓝及Ⅱ型胶原染色阳性;对照组明显变形,未形成软骨结构。结论单例先天性小耳残耳软骨细胞经体外扩增和动态诱导培养可在体外构建正常人大小耳廓软骨。
[Abstract]:Objective to investigate the feasibility of constructing normal human auricle tissue engineered cartilage in vitro by a single case of congenital microauricular residual ear chondrocytes. Methods A total of 40 patients with microauricular malformation were isolated. The cell proliferation was detected by MTT method and the amplification efficiency was calculated. The phenotype of cartilage of different algebraic cells was detected by immunofluorescence and PCR. The residual auricular chondrocytes of 3 patients were cultured to P3~P4 generation, and alginate gel was embedded in the auricle of normal human auricle, which was inoculated with poly (glycolic acid) / poly (lactic acid) scaffold. Histological staining was performed after 10 weeks of dynamic culture of chondrogenic cells in vitro. Results the extractable rate of chondrocytes was 3.90 卤1.27 脳 10 6 / g, and the proliferation ability of the cells was significantly increased in the proliferating medium, and the expression of proteoglycan was significantly decreased and the expression of type 鈪，

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