【作者】 毛英杰； 谷志远； 赵鹃；

【Author】 MAO Ying-jie,GU Zhi-yuan,ZHAO Juan (The Affiliated Stomatology Hospital,College of Medical Sciences,Zhejiang University,Hangzhou 310006,China)

【机构】 浙江大学医学院附属口腔医院修复科；

【摘要】 目的:研究不同细胞接种密度及饥饿对小鼠单核细胞RAW264.7诱导分化的影响。方法:RAW264.7细胞按5000/孔、1万/孔、2万/孔及4万/孔接种到48孔板,饥饿组饥饿12小时后用50ng/ml RANKL诱导RAW264.7细胞四天后,用抗酒石酸酸性磷酸酶(TRAP)染色法观察TRAP阳性多核细胞,用倒置显微镜观察多核细胞形态,非饥饿组正常培养12小时后用RANKL诱导观察。结果:光镜观察TRAP阳性多核破骨细胞(≥3个核)并计数,5000/孔饥饿组阳性多核细胞最多,非饥饿相对减少;1万/孔及2万/孔饥饿组较5000/孔饥饿组阳性多核细胞相对减少,但明显多于非饥饿组;4万/孔饥饿、非饥饿组少见阳性多核细胞。结论:RAW264.7细胞接种密度越低,50ng/ml RANKL诱导效果越好;RAW264.7细胞饥饿12小时后,50ng/ml RANKL诱导效果优于非饥饿组更多还原

【Abstract】 Objective:To study the effect of Initial Cells Seeding Density and Starvation on the differentiation of RAW264.7 cells induced by RANKL.Methods:RAW264.7 cells were seeded to 48 well paltes by the Cells Seeding Density of 5000、10000、20000 and 40000 per hole.RAW264.7 cells,starved for 12 hours or not,were treated with RANKL for 4 days.The morphological change was observed after TRAP staining and by inverted microscope.Results;It was lounil the most multinucleated osteoclast of positive TRAP staining was in the group of RAW264.7 cells,seeded by the Cells Seeding Density of 5000 per hole and starved for 12 hours.And, there were only a few multinueleated osteoclast of positive TRAP staining in the group of RAW264.7 cells,seeded by the Cells Seeding Density of 400000 per hole and starved for 12 hours or not.Conclusions:Initial Cells Seeding Density of 5000 per hole and Starvation is recommended for the induction of RAW264.7 cells hy RANKL.更多还原