新生兔气管软骨细胞的生物学特性

发布时间：2018-11-06 12:54

【摘要】：目的:探讨体外分离培养的新生兔气管软骨细胞的生物学特性。方法:通过酶消化法体外分离培养新生兔气管软骨细胞;倒置显微镜观察软骨细胞形态及生长状况;电镜观察软骨细胞超微结构;运用real-time PCR、免疫细胞化学染色和甲苯胺蓝染色检测气管软骨细胞分泌的细胞外基质成分。结果:体外分离、培养的兔气管软骨细胞呈短小三角形或不规则形贴壁生长。超微结构显示细胞较多突起,孔隙较多,胞质丰富,细胞器发达,细胞内可见大量蛋白分泌物。软骨细胞表达I、II型胶原、蛋白聚糖等,以II型胶原和蛋白聚糖表达为主。免疫细胞化学染色II型胶原和SOX9阳性,I型胶原弱阳性。甲苯胺蓝染色阳性。结论:适宜的酶消化单层培养法获得的新生兔气管软骨细胞具有分泌软骨细胞外基质成分的特性,可初步为体外构建组织工程气管治疗新生兔气管狭窄的实验研究提供种子细胞。
[Abstract]:Objective: to investigate the biological characteristics of trachea chondrocytes isolated from neonatal rabbits in vitro. Methods: neonatal rabbit trachea chondrocytes were isolated and cultured by enzyme digestion in vitro, the morphology and growth of chondrocytes were observed by inverted microscope, the ultrastructure of chondrocytes were observed by electron microscope. The extracellular matrix secreted by tracheal chondrocytes was detected by real-time PCR, immunocytochemical staining and toluidine blue staining. Results: cultured rabbit trachea chondrocytes grew in short triangle or irregular shape. The ultrastructure showed that the cells had more protuberances, more pores, abundant cytoplasm, well developed organelles, and a large number of protein secretions could be seen in the cells. Chondrocytes expressed type II collagen and proteoglycan, mainly II type collagen and proteoglycan. Immunocytochemical staining showed that II type collagen and SOX9 were positive, and type I collagen was weakly positive. Toluidine blue staining was positive. Conclusion: the new rabbit tracheal chondrocytes obtained by enzyme digestion monolayer culture have the characteristic of secreting extracellular matrix of chondrocytes. It may provide seed cells for the experimental study of trachea stenosis in newborn rabbits with tissue engineering trachea in vitro.
【作者单位】：广东省人民医院儿科;广东省医学科学院;广东省心血管病研究所小儿心脏外科;
【基金】：“十二五”国家科技支撑计划(No.2011BAI11B22)
【分类号】：R329.21

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