Trypsin is a very common enzyme used in cell culture to harvest cells by cleaving the proteins responsible for cell adhesion. However, trypsin also induces undesirable effects on cells, such as altering membrane proteins and the cytoskeleton, changing the composition of the cytoplasm and the cell volume, and even leading to cell death when used improperly. Using attenuated total reflection in the terahertz domain, confocal microscopy, and the propidium iodide test, we quantified in real time the change in cytoplasmic content induced by trypsin proteolysis on Madin-Darby canine kidney epithelial cells. We have observed a cytoplasmic modification from the very first seconds of trypsinization, following the change of cell volume due to mechanical re-equilibrium of the membrane. We found that the cytoplasmic alteration is associated with a transfer of small solutes: electrolytes and metabolites. We also found a very good nonlinear correlation between the side effects monitored by terahertz sensing and the cell height, regardless of the dependence of the cell height on trypsin concentration and exposure time.

中文翻译：

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太赫兹传感研究细胞分离过程中胰蛋白酶对细胞质的影响


胰蛋白酶是细胞培养中非常常见的酶，通过裂解负责细胞粘附的蛋白质来收获细胞。然而，胰蛋白酶也会对细胞产生不良影响，例如改变膜蛋白和细胞骨架，改变细胞质的组成和细胞体积，如果使用不当甚至导致细胞死亡。利用太赫兹域的衰减全反射、共聚焦显微镜和碘化丙啶测试，我们实时量化了胰蛋白酶水解对 Madin-Darby 犬肾上皮细胞引起的细胞质含量的变化。我们观察到，从胰蛋白酶化的第一秒起，随着膜的机械重新平衡导致细胞体积发生变化，细胞质发生了变化。我们发现细胞质的改变与小溶质的转移有关：电解质和代谢物。我们还发现太赫兹传感监测的副作用与细胞高度之间存在非常好的非线性相关性，无论细胞高度对胰蛋白酶浓度和暴露时间的依赖性如何。