CO_2及温度对显微活细胞培养系统动态观察活细胞Ca^(2+)变化的影响

Effects of CO_2 and temperature modulation on dynamic change of Ca^(2+) of living cells in a microscopic cell culture system

目的观察CO2及温度改变对显微活细胞培养系统动态观察活细胞Ca2+变化的影响。方法建立显微活细胞培养系统,CO2及温度可以通过CTI-Controller 3700 digital和Tempcontrol 37-2 digital进行控制改变,细胞分为A组:5%CO2维持37℃温度正常实验组、B组:无CO2但维持37℃温度组和C组:5%CO2室温温度组三个组,动态观察活细胞Ca2+荧光变化结果,并对结果进行比较分析。结果A组荧光强度从2 min到5 min逐步增强(P<0.05);B组荧光强度从1 min到5 min逐步减弱(P<0.05);C组荧光强度从2 min到5 min逐步减弱(P<0.05)。与A组的比较,B组从1 min到5 min时间点荧光强度分别减弱(P<0.05);与A组的比较,C组2 min到5 min时间点荧光强度分别减弱(P<0.05)。结论CO2及温度改变对动态观察活细胞Ca2+有较大的影响,维持5%CO2及37℃为活细胞动态观察实验中CO2及温度的最佳条件。

Objective To observe the effects of CO2 and temperature modulation on dynamic changes of Ca2＋ of living cells in a microscopic cell culture system.Methods A microscopic cell culture system was established.The modulation of CO2 and temperature was controlled by the CTI-Controller 3700 and Tempcontrol 37-2 digital equipments.Cells were divided into three groups： group A,control group with 5% CO2 and temperature 37℃;group B,experimental group with temperature 37℃ but no CO2;group C,experimental group with 5% CO2 and room temperature.After incubation,Ca2 ＋ fluorescence in living cells was dynamically observed and the results were analyzed.Results In group A,the fluorescence intensity gradually enhanced from 2 minutes to 5 minutes（P0.05）;in group B,fluorescence intensity gradually weakened from 1 minute to 5 minutes（P0.05）;in group C,fluorescence intensity gradually weakened from 2 minutes to 5 minutes（P0.05）.Compared with group A,the fluorescence intensity of group B weakened from 1 minute to 5 minutes（P0.05）;the fluorescence intensity of group C diminished from 2 minutes to 5 minutes（P 0.05）.Conclusions The changes of CO2 and temperature have great effect on the dynamic change of Ca2＋ in living cells.The optimal conditions for this experiment are 5% CO2 and 37 ℃.