软骨细胞特殊染色技术的比较与应用
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国家自然科学基金面上项目(项目编号:81272032);深圳市科工贸与信息委员会重点项目(项目编号:201101001);深圳市科创委资助项目(项目编号:JCY20130402114702130)。


Comparison of different special staining techniques of chondrocytes and their application values
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    摘要:

    目的 探讨多种特殊染色技术在软骨细胞中的染色规律及其应用价值。方法 选取出生7 d内的C57BL/6J小鼠12只, 处死后取双侧膝关节软骨行软骨细胞原代培养, 并行II型胶原免疫荧光鉴定, 取5代以内的原代细胞制作细胞爬片, 细胞爬片制备完成后行HE染色、番红O-固绿双染色、SA-β半染糖苷酶半乳糖苷酶衰老染色及II型胶原免疫组化染色, 观察软骨细胞组织形态变化, 并对几种染色方式进行比较。结果 HE染色中细胞核呈紫蓝色, 细胞质呈粉红色, 细胞形态结构清楚;番红O-固绿双染色中细胞核呈粉红色, 细胞质呈蓝绿色, 细胞形态结果较清晰;SA-β半染糖苷酶半乳糖苷酶衰老染色时, 衰老软骨细胞呈绿色, 未衰老细胞无着色;II型胶原免疫组化染色时, II型胶原呈棕黄色, 细胞核呈紫蓝色。结论 HE染色和番红O-固绿双染色可以清晰分辨细胞核和细胞质等细胞结构, 对单个细胞形态的显示比较清楚;SA-β半染糖苷酶半乳糖苷酶衰老染色可以显示出衰老细胞的数量及衰老的程度;II型胶原免疫组化染色可以对II型胶原进行定位与定量。

    Abstract:

    Objective To compare the advantages and values of several special staining methods of chondrocytes.Methods Twelve 7-day old healthy C57BL/6J mice were killed to obtain the cartilage tissue of the knee joint in order to isolate the chondrycytes. Type II collagen was used to assess the chondrocytes. Then the chondrocyte climbing slices were prepared. The materials were fixed, and HE staining, Safranin O-fast green staining, SA-β-gal staining and immunohistochemical staining of Type II collagen were performed and compared. Results HE staining showed clear morphology of the chondrocytes. The cell nuclei were stained blue and the cytoplasm was pink. Safranin O-fast green staining showed that the nuclei were pink and the cytoplasm green. SA-β-gal staining showed that the aging cells were green while the young cells were colorless. Immunohistochemical staining of type II collagen showed the distribution of type II collagen and they were stained brown while the cell nuclei were blue.Conclusions HE staining and safranin O-fast green staining can provide more information than the other staining techniques. SA-β-gal staining is useful in the analysis of aging chondrocytes. Immunohistochemical of type II collagen can be used to study type II collagen.

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于斐,曾晖,于红燕,雷鸣,袁昊,肖德明.软骨细胞特殊染色技术的比较与应用[J].中国比较医学杂志,2015,25(8):58~61,75.

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  • 最后修改日期:2015-07-12
  • 在线发布日期: 2015-09-01
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