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中国仓鼠V_(79)原代细胞在F_(12)培养基中培养(37℃、潮湿、5%CO_2、95%空气、pH=7.3),取指数生长细胞经处理置于T75瓶(2.5×10~5细胞/瓶),37℃培养12h,细胞于Lauda WGW/B水浴中作加热处理,温度变化小于0.05℃;经胰蛋白酶处理、冲洗、计数、形成集落,培养7~9天后固定、染色、测存活率,部分细胞用流式细胞光度计测细胞周期位置.高剂量率照射用15MeV线性加速器,5Gy/min,热处理后立即照射,低剂量率照射用~(60)Co,0.8Gy/min,与加热同时进行,源距细胞180cm.
Chinese hamster V_(79) primary cells were cultured in F_(12) medium (37°C, humid, 5% CO_2, 95% air, pH=7.3), exponentially growing cells were treated and placed in T75 bottles (2.5× 10~5 cells/bottle), cultured at 37°C for 12h, cells were heated in a Lauda WGW/B water bath, the temperature change was less than 0.05°C, treated with trypsin, washed, counted, formed colonies, cultured for 7 to 9 days, fixed, Staining, measurement of survival rate, some cells were measured by flow cytometry cell cycle location. High dose rate irradiation with 15MeV linear accelerator, 5Gy/min, immediately after heat treatment, low dose rate irradiation with ~ (60) Co, 0.8Gy /min, simultaneous with heating, source cell 180 cm.