CAJ | 학술논문

目的：建立新生SD大鼠髁突软骨下骨成骨细胞培养模型。方法用新生24 h SD大鼠，分离获取髁突，去尽软骨层，获得纯尽软骨下骨。采用改良贴壁组织块反复消化法培养细胞。通过相差显微镜和HE染色观察其细胞形态，并且用碱性磷酸酶染色和钙化结节染色方法进行细胞鉴定，噻唑蓝（MTT）比色法检测其增值能力。结果细胞呈多种形态，有三角形、梭形、不规则形。碱性磷酸酶染色和钙化结节染色均呈阳性，细胞接种后1～3 d内细胞增殖缓慢，第8天达到高峰，以后增长速度逐渐减慢。结论该方法获得新生SD大鼠髁突软骨下骨成骨细胞能在体外稳定培养，并且细胞具有典型的成骨细胞形态和功能。
목적：건립신생SD대서과돌연골하골성골세포배양모형。방법용신생24 h SD대서，분리획취과돌，거진연골층，획득순진연골하골。채용개량첩벽조직괴반복소화법배양세포。통과상차현미경화HE염색관찰기세포형태，병차용감성린산매염색화개화결절염색방법진행세포감정，새서람（MTT）비색법검측기증치능력。결과세포정다충형태，유삼각형、사형、불규칙형。감성린산매염색화개화결절염색균정양성，세포접충후1～3 d내세포증식완만，제8천체도고봉，이후증장속도축점감만。결론해방법획득신생SD대서과돌연골하골성골세포능재체외은정배양，병차세포구유전형적성골세포형태화공능。
Objective Establish an experimental model for primary subchondral bone osteoblasts culture of neonatal SD rat's condylar process. Methods Under the condition of sterile,24-hour SD rat was executed and its condylar process was isolated. Removing cartilage layer, the subchondral bone was exposed obviously, then it was cultured with modified repeating enzymatic digestion-adherent explants method. The cellular morphology was identified with invert microscope and immunohistochemistry staining, the osteoblasts were identified by alkaline phosphorase (ALP) staining and calcified nodules staining, and the proliferation of the acquired cells was examined by methyl thiazolyl tetrazolium (MTT) assay. Results A variety of cell morphologies were observed, such as spindle-shaped, triangular and irregular-shape, and their cell processes were significant. The alkaline phosphatase staining and calcified nodules staining of cultured osteoblasts with mineralized nodules were positive. Cells grew slowly in 1-3 days, and the cells growth reached the highest level at the 8th day. The cells growth trend has gradually slowed down after 8 days. Conclusion The method is an efficient way to culture and obtain purified neonatal SD rat's subchondral bone osteoblasts with typical characteristics.