摘要
目的设计、组装开放式压力控制培养系统,研究该系统对培养液Ph值、碳酸氢根（HCO3-）浓度、氧气分压（Po2）、二氧化碳分压（Pco2）及视网膜色素上皮细胞增殖的影响,评价该系统在青光眼基础研究中应用的可行性.方法采用动态循环气体控制压力的方法,设计组装开放式压力控制培养系统.比较正常压力细胞培养系统（37℃恒温培养箱,5%CO2浓度）、开放式压力控制培养系统、密闭压力控制培养系统对DMEM/F-12培养液Ph值、HCO3-浓度、Po2、Pco2的影响.采用MTF法比较3种培养系统对人视网膜色素上皮细胞增殖能力的影响.结果成功组装开放式压力控制培养系统,通过调节进出气流速度,可将培养瓶内压力控制在0～100 mm Hg（1 mm Hg=0.133kPa）之间.培养24 h测定结果：开放式与正常压力细胞培养系统组比较,其Ph值（P=0.927）、HCO3-浓度（P=0.887）、Po2（P=0.818）、Pco2（P=0.770）、A值（P=0.719）差异均无统计学意义;密闭组与正常压力细胞培养系统组比较,前者Ph值升高（P=0.001）,HCO3-浓度（P=0.000）,Po2（P=0.000）、Pco2（P=0.000）、A值（P=0.000）降低.结论开放式压力控制培养系统设计、组装成功,可以实现对培养环境压力及培养液Ph值、HCO3-浓度、Po2及Pco2的均衡调控.相对于密闭压力控制培养系统,该系统可以排除培养条件差异对研究结果的影响.标准化的开放式压力控制培养系统有望为青光眼体外研究提供理想实验模型.
Objective To develop and set up a new culture system,which can apply pressure to cultured cells with open cycling air. The effects of this new system on the pH value, HCO3^-concentration, O2 pressure （pO2）, CO2 pressure （pCO2）and the proliferation of retinal pigment epithelium （RPE） were tested to evaluate its efficiency in the study of glaucoma. Methods In the open cycling air pressure control culture system, the pressure inside the culture flasks was controlled by increase or decrease of the perfuse airflow. The influence of different culture systems （normal pressure culture system,open cycling air pressure control system and occlusive pressure control system） on the pH value, HCO3^- concentration, pO2, pCO2 and proliferation of RPE were tested. The data were analyzed with SPSS software. Results The open cycling air pressure control culture system worked effectively, the pressure inside the culture flask can be controlled from 0 to 100 mm Hg. The difference of pH value, HCOf concentration, pO2, and pCO2 of culture medium and the proliferation of RPE between normal pressure culture system and open cycling air pressure control system were not significant （ P = 0. 927,0. 887,0. 818,0. 770,0. 719, respectively）. There was significant difference in these data between normal pressure culture system and occlusive pressure control system （ P = 0. 001, 0. 000,0. 000,0. 000,0. 000,respectively）. Conclusions A new designed standard culture system applying pressure to cells with open cycling air was effective at pressure controlling and pH value, HCO3^- concentration,pO2 and pCO2 controlling. This system may act as an ideal model in the experimental study of glaucoma. （Chin J Ophtbalmol , 2006,42 ： 513-516）
出处
《中华眼科杂志》
CAS
CSCD
北大核心
2006年第6期513-516,共4页
Chinese Journal of Ophthalmology
