小鼠鼻软骨细胞的培养

Culture of mouse nasal chondrocyte

目的 研究小鼠鼻软骨细胞体外培养方法及形态特征。方法 取Balb/c小鼠子鼠的鼻中隔软骨 ,随机分为 4组后分别消化 4h ,8h ,12h及 16h ,观察体外培养软骨细胞的生长状况 ,并用Ⅱ型胶原抗体进行细胞来源鉴定。结果 经Ⅱ型胶原酶孵育消化 4h的组织 ,细胞量少 ,生长缓慢 ,无法进行正常传代。而消化 8h的软骨组织、细胞于 1d后即开始贴壁、伸展 ,9d后第一次传代 ,传代细胞生长稳定 ,形态多样 ,Ⅱ型胶原抗体染色阳性。消化 2h及 16h的细胞几乎不贴壁。结论 采用酶消化法可获得体外培养的小鼠鼻软骨细胞 ,消化时间以 8h为佳。

Objective To study the method for culturing nasal chondrocytes and the morphological characters of these cells. Methods Nasal septal cartilages of Balb/c mouse were separated and divided into four groups. Cartilages were digested with typeⅡcollagenase for 4 hrs,8 hrs,12 hrs and 16 hrs respectively, then cultured. Cell morphologies were observed and their origin were identified with typeⅡcollagen antibody. Results Cells collected from cartilage tissues digested for 4 hrs were few and couldn′t be generated; While cells from those digested for 12hrs and 16hrs were almost all dead and didn't stick to the walls of the culture bottles. However, cells from tissues digested for 8 hrs began to stick to the walls and extended after 1d incubated and generated for the first time after 9 d. The generated cells appeared different shapes and expressed typeⅡcollagen protein. Conclusion Mouse nasal chondrocyte's can be obtained by digesting with typeⅡcollagenase. In addition, 8 hrs was the optimized time for digesting in this in vitro system.