用于细胞培养实验的定量抽气-充气方法及相配装置  

A Method of Exhausting and Gassing in a Quantitative Manner and Related Apparatuses for Cell Culture Experiments

作　　者：潘轩[1] 李璟 熊蕾[1] 唐文岘[1] 张君[1] 汪保和[1,3] 

机构地区：[1]湖南师范大学生命科学学院,湖南长沙410081 [2]湖南省老年医院,湖南长沙410016 [3]湖南省生物发育工程及新产品研发协同创新中心,湖南长沙410081

出　　处：《生物技术通讯》2015年第5期691-696,共6页Letters in Biotechnology

基　　金：湖南省科技计划重点项目(2014TT1008);湖南省生态学重点学科基金(0713)

摘　　要：目的:建立一种用于细胞培养实验的定量抽气-充气方法,研制与该方法配用的装置。方法:推导用于定量抽气和充气的一系列公式,设计和制作抽气-充气装置和配气-输气装置;用这2种装置配制不同组分配比的混合气体,检测O2和CO2浓度;进行细胞培养实验,验证所述方法和装置的实用性。结果:在不同组分配比的混合气体中,O2体积分数检测值与预定值的差值均<1.0%,CO2体积分数检测值与预定值的差值均<1.2%。在1%、20%和80%O2浓度培养条件下,每培养瓶MC-3T3-E1细胞的计数分别为(96.5±29.6)×103、(250.4±81.0)×103和(8.8±3.9)×103;与20%O2浓度条件比,细胞数差异有统计学显著性。在1%、20%和40%O2浓度培养条件下,每106小鼠骨髓有核细胞的CFU-F集落生成数分别为8.0±3.3、17.7±4.7和2.3±0.8;与20%O2浓度培养条件比,低氧和高氧条件下的CFU-F集落形成率均显著性减低。结论:定量抽气-充气方法及相配装置的可靠性高、适用性强、费用低廉,可作为细胞培养气相环境建立和维持方法和装置的有益选择。Objective： To develop a method of exhausting and gassing in a quantitative manner and related appa- ratuses for ceil culture experiments. Methods： A group of equations were derived for the quantitative calculation of the exhausting and gassing. The gas exhausting-gassing device and the gas preparing-delivering device were de- signed and fabricated. The gas mixtures of predetermined proportions were prepared using these devices, and the concentrations of 02 and CO2 in the gas mixtures were measured. Cell cuhure experiments were performed to veri- fy the practicability of the described method and apparatuses. Results： In the gas mixtures of various proportions, the differences between measured and predetermined values of oxygen volume fraction were lower than 1.0%, and the differences between measured and predetermined values of carbon dioxide volume fraction was lower than 1.2%. The counts of MC-3T3-E1 cells per culture flask were （96.5±29.6）×10^3, （250.4±81.0）×10^3 and （8.8±3.9）× 10^3 respectively at oxygen concentration of 1, 20 and 80 percent; and the intergroup differences of cell numbers were statistically significant compared with the 20 percent oxygen concentration. The yields of CFU-F colonies were 8.0±3.3, 17.7±4.7 and 2.3±0.8 per 10^6 nucleated cells from murine bone marrow under oxygen tensions of 1, 20 and 40 percent, respectively. CFU-F efficiencies at lower or higher levels of oxygen were decreased significantly in comparison with those at 20 percent oxygen. Conclusion： Our method of exhausting and gassing in a quanti- tative manner and related apparatuses are very reliable, widely applicable and cost-saving, which could be consid- ered as useful alternatives to the means for establishing and maintaining the gaseous atmosphere of cell cultivation.

关 键 词：细胞培养 气体混配 气体输送 方法 装置 

分 类 号：Q813.1[生物学—生物工程]