【摘要】 目的 了解体外培养成骨细胞的接种数量对钙化结节形成的影响。方法 实验选用了1×105个/ml、2×105个/ml、5×105个/ml三个不同的细胞接种浓度，并且每个浓度组分别给予含β-甘油磷酸钠和维生素C的条件培养液以及常规培养液进行细胞培养。显微镜观察钙化结节的形成，并做钙化结节染色。结果 在培养30天内，条件培养液或常规培养液培养的5×105个/ml组细胞无结节形成；而2×105个/ml组和1×105个/ml组都有钙化结节形成，而且有细胞越少形成结节越早，数量越多的趋势；给予条件培养液又较常规培养液培养有结节形成早而多的趋势。钙化结节染色显示结节为红色。结论 体外培养成骨细胞钙化结节的形成须接种适宜的细胞数量，用条件培养液培养细胞能促进钙化结节形成。

    【关键词】 成骨细胞； 钙化结节； 体外培养

    Observation of cultured osteoblasts forming mineralization nodes in vitro

    YIN Mei-zhen, LI Shi-pu, CAO Xian-ying，et al.

    Biomedical Material and Engeneering Center, Wuhan University of Technology, Wuhan 430070,China

    【Abstract】 Objective To investigate whether osteoblasts cultured in vitro can form mineralization nodes. Method The osteoblasts at 1×105/ml,2×105/ml and 5×105 /ml cell density were cultured in the DMEM medium and in the DMEM medium supplemented with β-glycerophosphate sodium and ascorbic acid. The formation process of mineralization nodes were observed with microscope and calcified nodules stain was made. Results There were more mineralization nodes to form in shorter time in the 1×105/ml cell density group compared with in the 2×105/ml cell density group, but in the 5×105/ml cell density group, there were not, and with the DMEM medium contained β-glycerophosphate sodium and ascorbic acid, mineralization nodes formed more early compared with the DMEM medium. Mineralization nodes stain showed red.Conclusion Cultured osteoblasts at suitable cell density can easily form mineralization nodes in vitro, and β-glycerophosphate sodium and ascorbic acid just promote formation of mineralization nodes. ......