软骨细胞爬片特殊染色技术的改进与应用中国试验动物学报.doc

软骨细胞特殊染色技术的比较与应用R332 【文献标识码】 【文章编号】 Comparison of Values in Different Special Staining Methods on Chondrocytes Morphology YU Fei1,ZENG Hui1,YU Hongyan2,LEI Ming1,YUAN Hao1,XIAO Deming1* (1Peking University Shenzhen Hospital,Guangdong Shenzhen 518036,China. 2Municipal Hospital of Binzhou,Shandong Binzhou,256600,China) 【Abstract】 Objective To compare the advantages and values of several different special staining methods in chondrocyte morphology. Methods 12 7-day old healthy C57BL/6J mice were killed to abtaion the cartilage tissue of the knee joint in order to get the chondrycytes. Type II collagen was used to appraisal the chondrocytes.Then use the chondrocytes to make cell climbing pieces. The materials were fixed.HE staining, Safranin O-Fast Green staining, SA-β-gal staining and immunohistochemical of Type II collagen were compared. Results HE staining could show the comprehensive of morphology in chondrocytes.The nucleus were blue and the cytoplasm were pink.Safranin O-Fast Green staining showed that the nucleus were pink and the cytoplasm were green. SA-β-gal staining showed that aging cells were green compared with the young cells were colorless Immunohistochemical of Type II collagen could show the distribution of Type II collagen and Type II were brown while the nucleus were blue. Conclusion HE staining and Safranin O-Fast Green staining can get more information than others. SA-β-gal staining is useful in the analysis of aging chondrocytes.Immunohistochemical of Type II collagen can be used to research Type II collagen. 【KEY WORDS】 chondrocytes,special staining,cell climbing pieces,primary cells,knee joint 骨关节炎发病机制及治疗措施的研究中经常用到细胞学水平的研究，观察软骨细胞的形态结构成为实验中的一种重要的方法。在过去的研究[1,2,3]中，人们对软骨大体标本的特殊染色技术已经进行了大量的改良，并应用于骨关节炎的研究中，取得了良好的效果，但是对于软骨细胞水平的染色技术发展缓慢，以至于无法用简便的方法对细胞形态进行观察，对研究造成了不便。本研究利用C57BL/6J小鼠进行软骨细胞的原代培养，对软骨细胞爬片进行HE染色、番红O-固绿双染色、SA-β半染糖苷酶衰老染色及II型胶原免疫组化染色，探讨这几种染色技术在软骨细胞爬片中的染色规律及应用价值，为骨关节炎细胞水平研究提供便利的条件。 1 材料和方法 1.1 实验材料 1.1.1 实验动物及实验环境 出生7天内的健康SPF级C