CAJ | 학술논문

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。
목적：탐토체외분리배양적신생토기관연골세포적생물학특성。방법：통과매소화법체외분리배양신생토기관연골세포；도치현미경관찰연골세포형태급생장상황；전경관찰연골세포초미결구；운용real-time PCR、면역세포화학염색화갑분알람염색검측기관연골세포분비적세포외기질성분。결과：체외분리、배양적토기관연골세포정단소삼각형혹불규칙형첩벽생장。초미결구현시세포교다돌기，공극교다，포질봉부，세포기발체，세포내가견대량단백분비물。연골세포표체I、II형효원、단백취당등，이II형효원화단백취당표체위주。면역세포화학염색II형효원화SOX9양성，I형효원약양성。갑분알람염색양성。결론：괄의적매소화단층배양법획득적신생토기관연골세포구유분비연골세포외기질성분적특성，가초보위체외구건조직공정기관치료신생토기관협착적실험연구제공충자세포。
[ ABSTRACT] AIM:To investigate the biological characteristics of newborn rabbit tracheal chondrocytes in vitro. METHODS:Newborn rabbit tracheal chondrocytes were obtained by the method of enzyme digestion, and then cultured in monolayer in vitro.Morphological and growth observations were performed under inverted phase contrast microscope.The ultrastructures of the cells were observed under scanning electron microscope and transmission electron microscope.The bi-ological characteristics of secreted extracellular matrix components were detected by real-time PCR, immunocytochemistry staining and toluidine blue staining.RESULTS: Newborn rabbit tracheal chondrocytes isolated and cultured in vitro showed short triangular or irregular shapes, and adherent growth very well.The ultrastructures of the cells showed pore and abundant cytoplasm and organelles, with a lot of protein secretions in the cells.The chondrocytes expressed the mRNA of collagen I, collagen II and proteoglycans, mainly collagen II and proteoglycans.Immunocytochemistry staining showed col-lagen II and SOX9 positive, and collagen I weakly positive.Toluidine blue staining was also positive.CONCLUSION:Enzyme digestion and monolayer culture are suitable method to obtain newborn rabbit tracheal chondrocytes.These cells, secreting extracellular matrix components, are able to be selected as seed cells for tissue engineering of trachea in vitro, and used to study the therapeutic method for neonatal rabbit tracheal stenosis.