人鼻中隔软骨细胞培养法的改良及其生物学特性观察

The modified method for human nasal septum chondrocyte culture and the study of biological characters of the chondrocyte

目的对人鼻中隔软骨细胞培养法进行改良并观察改良法培养的人鼻中隔软骨细胞的生物学特性。方法采用软骨细胞悬液和软骨块共同培养法体外培养人鼻中隔软骨细胞,对其进行倒置相差显微镜动态观察、甲苯胺蓝异染和Ⅱ型胶原的免疫组织化学染色,并测定35S-Na2SO4掺入量以观察软骨细胞蛋白多糖合成量,在细胞表型和功能方面进行鉴定并与单纯细胞悬液培养法比较。结果改良法获得的人鼻中隔软骨细胞存活率高,量更多,且细胞的生长过程、特殊染色无明显差别,35S-Na2SO4掺入量与常规法差异无显著性意义(P>0.05)。结论软骨细胞悬液和软骨块共同培养法是一种可行的细胞培养法,培养的人鼻中隔软骨细胞生物特性良好。

OBJECTIVE To established a viable and convenient method for human nasoseptal chondrocyte culture in vitro and study the biological characters of the chondrocyte. METHODS The chondrocytas were obtained by incomplete enzyme digestion of cartilage and were seeded with small piece of cartilage in 25 ml culture flasks coated with polylysine previously. This study determines the biologic synthesis of the chondrocytes by toluidine blue and type Ⅱ collagen immunohistochemistry reaction. The proteoglycan synthesis efficiencies of human nasoseptal chondrocytes cultured in vitro were measured by using ^36S-Na2SO4 incorporation. The modified method and the conventional method in proteoglycan synthesis were also compared. RESULTS The results suggested that human nasoseptal chondrocytes cultured by modified method still show the normal structure and morphology, and maintain the function of metabolism and self duplication. With the modified method, we can get a higher cell survival rate and mere cells. CONCLUSION The modified method is a viable and convenient method.