非CO2依赖型细胞培养系统对膀胱癌原代细胞增殖的影响-泌尿外科专业论文.docxVIP

PAGE PAGE 5 37℃ (大气环境) 培养箱条件下细胞克隆数（66±6）比 CO2 依赖型细胞培 养系统、37℃（5%，CO2）培养箱条件下明显减少（P0.05）。 3、细胞在非 CO2 依赖型细胞培养系统、37℃ (大气环境) 培养箱与 CO2 依赖型细胞培养系统、37℃（5%，CO2）培养箱条件下培养，细胞生长增殖 趋势基本一致（P0.05）,；而在 CO2 依赖型细胞培养系统、37℃ (大气环 境) 培养箱条件下，比 CO2 依赖型细胞培养系统、37℃（5%，CO2）培养箱条 件下生长增殖显著下降（P0.05）。 4、细胞在非 CO2 依赖型细胞培养系统、37℃ (大气环境) 培养箱与 CO2 依赖型细胞培养系统、37℃（5%，CO2）培养箱条件下培养，随着作用时间 的延长细胞凋亡数增加，凋亡率相比无明显变化（P0.05）；而在 CO2 依赖 型细胞培养系统、37℃ (大气环境) 培养箱条件下比 CO2 依赖型细胞培养系 统、37℃（5%，CO2）培养箱条件下细胞凋亡数明显增加（P0.05）。 5、细胞在非 CO2 依赖型细胞培养系统、37℃ (大气环境) 培养箱与 CO2 依赖型细胞培养系统、37℃（5%，CO2）培养箱条件下培养，表达 survivin、 caspase-3 因子与蛋白无明显变化（P0.05）；而在 CO2 依赖型细胞培养系统、 37℃ (大气环境) 培养箱条件下随着培养时间的延长，caspase-3 因子与蛋 白表达增高， survivin mRNA 及其蛋白的表达并未明显变化，比 CO2 依赖型 细胞培养系统、37℃（5%，CO2）培养箱条件下表达上调（P0.05）。 结论 膀胱癌非 CO2 依赖型细胞培养系统对细胞增殖、基因表达等方面 与膀胱癌 CO2 依赖型细胞培养系统无明显区别。 关键词 膀胱癌原代细胞；非 CO2 依赖；survivin；caspase-3 The Effects of CO2-independent Cell Culture System on The Proliferation of Primary Bladder Cancer Cells ABSTRACT Objective To evaluate the effects of CO2-independent culture cell system on the prolilferation of primary bladder cancer cells .Whether there are significant differences in CO2-dependent and CO2-independent cell culture systems. Methods primary bladder cancer cells culture using CO2-independent culture cell system at 37℃ in 5% carbon dioxide incubator and 37℃ in air incubator,Meamwhile using CO2-dependent culture cell system at 37℃ in 5% carbon dioxide incubator and 37℃ in air incubator. The testing of pH value 7 days in the two culture cell systems.Differences of the cell viability were assayed with MTS test and Colony forming assay. The cell cycle and apoptosis ratio of primary bladder cancer cells were treated in the two culture cell systems throuth FCM.The expression of survivin and caspase-3 mRNA of cell culturing with two system above detected by RT-PCR. The expression of aurvivin and caspase-3 protein of cell culturing with two system above detected by Western bolt. Results 1. Culturing with CO2-independent culture cell system at 37℃