软骨细胞膜片的成软骨性能
发布时间：2018-07-27 10:36
【摘要】：背景:基于温敏培养皿的细胞膜片技术在组织工程研究中受到越来越多的关注,但是温敏培养皿价格高昂,有必要研究替代方法。目的:应用普通培养皿进行细胞膜片培养,并分析软骨细胞膜片的成软骨性能。方法:取1例鼻中隔偏曲患者手术切除的鼻中隔软骨消化提取原代软骨细胞,扩增培养。取第3代细胞在体外密集培养,使之分泌细胞外基质,形成单层细胞膜片;另取第2代软骨细胞,叠加接种于单层细胞膜片上,使其贴覆生长,形成双层细胞膜片。应用细胞刮刀收获细胞膜片,研究其性状,进一步在体外培养、植入裸鼠体内构建出人工软骨块。结果与结论:(1)单层细胞膜片和双层细胞膜片均形成质软的凝胶样结构,大体观察两者无明显区别;(2)新刮起的细胞膜片具有良好的可流动性和成胶性;植入裸鼠体内培育8周之后,均形成成熟的软骨块;(3)组织学观察见细胞膜片是由成层排列的软骨细胞和细胞外基质组成的薄片,在体外培养过程中即有糖胺聚糖和Ⅱ型胶原的初步表达,植入体内8周之后形成的软骨呈现为成熟的软骨组织;(4)生化定量检测结果表明,DNA含量高于天然鼻中隔软骨,而糖胺聚糖和羟基脯氨酸含量与天然软骨差异无显著性意义;(5)结果提示,应用普通培养皿可成功构建并获取细胞膜片,细胞膜片呈现良好的成软骨性能。
[Abstract]:Background: cell membrane technology based on thermo-sensitive petri dish has attracted more and more attention in tissue engineering, but the price of thermo-sensitive dish is high, so it is necessary to study alternative methods. Aim: to study the chondrogenic properties of chondrocytes in vitro. Methods: the primary chondrocytes were extracted from a patient with nasal septum deviation. The third generation of chondrocytes were cultured in vitro to secrete extracellular matrix to form monolayer cell membrane, and the second generation chondrocytes were inoculated on the monolayer cell membrane to form double layer cell membrane. The cell scraper was used to harvest the cell membrane and to study its properties. The artificial chondrocytes were further cultured in vitro and implanted into nude mice to construct artificial chondrocytes. Results and conclusions: (1) soft gelatin-like structures were formed in monolayer and bilayer cell membranes, and there was no significant difference between them, (2) the newly scraped membranes had good fluidity and gelling ability. After 8 weeks of implantation, mature chondrocytes were formed in nude mice. (3) histological observation showed that the membrane slices were composed of chondrocytes and extracellular matrix. The primary expression of glycosaminoglycan and type 鈪，
本文编号：2147552
[Abstract]:Background: cell membrane technology based on thermo-sensitive petri dish has attracted more and more attention in tissue engineering, but the price of thermo-sensitive dish is high, so it is necessary to study alternative methods. Aim: to study the chondrogenic properties of chondrocytes in vitro. Methods: the primary chondrocytes were extracted from a patient with nasal septum deviation. The third generation of chondrocytes were cultured in vitro to secrete extracellular matrix to form monolayer cell membrane, and the second generation chondrocytes were inoculated on the monolayer cell membrane to form double layer cell membrane. The cell scraper was used to harvest the cell membrane and to study its properties. The artificial chondrocytes were further cultured in vitro and implanted into nude mice to construct artificial chondrocytes. Results and conclusions: (1) soft gelatin-like structures were formed in monolayer and bilayer cell membranes, and there was no significant difference between them, (2) the newly scraped membranes had good fluidity and gelling ability. After 8 weeks of implantation, mature chondrocytes were formed in nude mice. (3) histological observation showed that the membrane slices were composed of chondrocytes and extracellular matrix. The primary expression of glycosaminoglycan and type 鈪，
本文编号：2147552
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