CAJ | 학술논문

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7d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。
목적：탐토다충특수염색기술재연골세포중적염색규률급기응용개치。방법선취출생7d내적C57BL／6J소서12지，처사후취쌍측슬관절연골행연골세포원대배양，병행II형효원면역형광감정，취5대이내적원대세포제작세포파편，세포파편제비완성후행HE염색、번홍O－고록쌍염색、SA－β반염당감매반유당감매쇠로염색급II형효원면역조화염색，관찰연골세포조직형태변화，병대궤충염색방식진행비교。결과 HE염색중세포핵정자람색，세포질정분홍색，세포형태결구청초；번홍O－고록쌍염색중세포핵정분홍색，세포질정람록색，세포형태결과교청석；SA－β반염당감매반유당감매쇠로염색시，쇠로연골세포정록색，미쇠로세포무착색；II형효원면역조화염색시，II형효원정종황색，세포핵정자람색。결론 HE염색화번홍O－고록쌍염색가이청석분변세포핵화세포질등세포결구，대단개세포형태적현시비교청초；SA－β반염당감매반유당감매쇠로염색가이현시출쇠로세포적수량급쇠로적정도；II형효원면역조화염색가이대II형효원진행정위여정량。
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 .