【摘要】 背景与目的放射生物学试验时单层细胞或移植瘤均存在不同程度的局限性,而多细胞球体(multicellularspheroid,MTS)作为实体瘤的模型应用较多。本研究旨在了解A549肺腺癌MTS的生长特性,有助于选择最佳MTS用于实验研究。方法1.75×107的A549细胞株35mL转移入2%琼脂糖封底的100mm培养皿中旋转培养,60r/min,40h时分离MTS到2%琼脂糖封底的96孔板中,每孔一个MTS,静止培养。结果旋转培养阶段A549MTS形态较圆,大小均一,细胞间连接不紧密。经过约10天的静止培养,直径在429~570μm时MTS的细胞间连接紧密,静止培养26.5天时直径达1358.5μm,已经不能用于试验。电镜观察MTS的细胞生长良好,很少有凋亡和坏死。MTS的细胞周期以G0~G1期占优势。结论旋转结合静止培养方法简单,所得MTS的生长特性与实体瘤相似,直径在500μm左右时最适合用于实验研究。更多还原

【Abstract】 Background and objective Owing to the limitation of monolayer tumour cells or transplanted tumor in radiation biology, multicellular spheroid (MTS) as a model of entity tumor had a wide range of application. This research aims to study the growth characteristics of A549 lung adenocarcinoma MTS so as to choose the best ones to perform experiments. Methods 1.75×10~7 A549 cells in 35mL were transfered into the 100mm incubator sealed with 2% agarose to have a rotating culture with 60r/min. After 40hours every MTS was transfered into each of the 96 wells of multi-well plates coated with 2% agarose to have a static culture. Results During the rotation stage the MTS was round, the size was uniform, and the conjunction between cells was loose. After about 10 days of static culture, when the diameter of MTS at 429 to 570μm the conjunction was close. After 26.5 days of static culture, the diameter of MTS became 1358.5μm which could not be used for experiment. Observed under electron microscopy the cells in the MTS grew well without any apoptosis and necrosis. The cell cycle of MTS was dominated by G0 and G1 stage. Conclusion Combination of rotating and static culture is a simple method to produce MTS which has similar growth characteristics with entity tumor and is best suitable for experiment research when the diameter is about 500μm.更多还原