Nikulin S V, Knyazev E N, Poloznikov A A, Shilin S A, Gazizov I N, Zakharova G S, Gerasimenko T N
Bioclinicum Research and Development Center, Moscow, 115088 Russia.
Moscow Institute of Physics and Technology (State University), Dolgoprudny, Moscow oblast, 141701 Russia.
Mol Biol (Mosk). 2018 Jul-Aug;52(4):667-674. doi: 10.1134/S0026898418040134.
Drug bioavailability studies commonly employ in vitro barrier tissue models consisting of epithelial and endothelial cells. These experiments require that the cell barrier quality be assessed regularly, which is usually performed using various labeled substrates and/or evaluation of transepithelial (transendothelial) electrical resistance (TEER). This technique provides information on the integrity of the monolayer, but not on differentiation-induced changes in the cell morphology. The present work shows that impedance spectroscopy can be applied to monitor both the integrity of the monolayer and the morphological changes of Caco-2 cells. The growth kinetics of the apical membrane was determined by calculating the electrical capacitance of the cell monolayer. In the course of differentiation, the most pronounced changes in the expression levels were observed for the mRNAs that encode SLC30A10 and SLC23A3 transporters. Their increase correlated with an increase in the apical membrane area, indicating that SLC30A10 and SLC23A3 mRNA levels assessed by qRT-PCR may be employed as cell differentiation biomarkers in Caco-2 models.
药物生物利用度研究通常采用由上皮细胞和内皮细胞组成的体外屏障组织模型。这些实验要求定期评估细胞屏障质量,这通常使用各种标记底物和/或跨上皮(跨内皮)电阻(TEER)评估来进行。该技术提供了关于单层完整性的信息,但没有提供关于分化诱导的细胞形态变化的信息。目前的工作表明,阻抗谱可用于监测单层的完整性和Caco-2细胞的形态变化。通过计算细胞单层的电容来确定顶端膜的生长动力学。在分化过程中,观察到编码SLC30A10和SLC23A3转运蛋白的mRNA表达水平变化最为明显。它们的增加与顶端膜面积的增加相关,表明通过qRT-PCR评估的SLC30A10和SLC23A3 mRNA水平可作为Caco-2模型中的细胞分化生物标志物。