1 BioMediTech Institute and Faculty of Biomedical Sciences and Engineering, Tampere University of Technology, Tampere, Finland.
2 BioMediTech Institute and Faculty of Medicine and Life Sciences, University of Tampere, Tampere, Finland.
SLAS Technol. 2018 Dec;23(6):566-579. doi: 10.1177/2472630318768710. Epub 2018 May 3.
A physiologically relevant environment is essential for successful long-term cell culturing in vitro. Precise control of temperature, one of the most crucial environmental parameters in cell cultures, increases the fidelity and repeatability of the experiments. Unfortunately, direct temperature measurement can interfere with the cultures or prevent imaging of the cells. Furthermore, the assessment of dynamic temperature variations in the cell culture area is challenging with the methods traditionally used for measuring temperature in cell culture systems. To overcome these challenges, we integrated a microscale cell culture environment together with live-cell imaging and a precise local temperature control that is based on an indirect measurement. The control method uses a remote temperature measurement and a mathematical model for estimating temperature at the desired area. The system maintained the temperature at 37±0.3 °C for more than 4 days. We also showed that the system precisely controls the culture temperature during temperature transients and compensates for the disturbance when changing the cell cultivation medium, and presented the portability of the heating system. Finally, we demonstrated a successful long-term culturing of human induced stem cell-derived beating cardiomyocytes, and analyzed their beating rates at different temperatures.
在体外成功进行长期细胞培养，生理相关的环境至关重要。精确控制温度是细胞培养中最重要的环境参数之一，可提高实验的准确性和可重复性。然而，直接的温度测量会干扰培养物或阻止细胞成像。此外，使用传统的细胞培养系统测温方法，评估细胞培养区域中的动态温度变化具有挑战性。为了克服这些挑战，我们将微尺度细胞培养环境与活细胞成像以及基于间接测量的精确局部温度控制集成在一起。该控制方法使用远程温度测量和数学模型来估算所需区域的温度。该系统可将温度维持在 37±0.3°C 超过 4 天。我们还表明，该系统可在温度瞬变期间精确控制培养物温度，并在更换细胞培养液时补偿干扰，同时展示了加热系统的便携性。最后，我们成功地对人诱导多能干细胞来源的搏动心肌细胞进行了长期培养，并在不同温度下分析了它们的搏动率。
