Kato Momoko, Sato Kae
Department of Chemical and Biological Sciences, Faculty of Science, Japan Women's University, 2-8-1 Mejirodai, Bunkyo, Tokyo 112-8681, Japan.
Micromachines (Basel). 2025 Jan 15;16(1):93. doi: 10.3390/mi16010093.
Microfluidic-based cell-stretching devices are vital for studying the molecular pathways involved in cellular responses to mechanobiological processes. Accurate evaluation of these responses requires detailed observation of cells cultured in this cell-stretching device. This study aimed to develop a method for preparing microscope slides to enable high-magnification imaging of cells in these devices. The key innovation is creating a peelable bond between the cell culture membrane and the upper channel, allowing for easy removal of the upper layer and precise cutting of the membrane for high-magnification microscopy. Using the fabricated device, OP9 cells (15,000 cells/channel) were stretched, and the effects of focal adhesion proteins and the intracellular distribution of YAP1 were examined under a fluorescence microscope with 100× and 60× objectives. Stretch stimulation increased integrinβ1 expression and promoted integrin-vinculin complex formation by approximately 1.4-fold in OP9 cells. Furthermore, YAP1 nuclear localization was significantly enhanced (approximately 1.3-fold) during stretching. This method offers a valuable tool for researchers using microfluidic-based cell-stretching devices. The advancement of imaging techniques in microdevice research is expected to further drive progress in mechanobiology research.
基于微流控的细胞拉伸装置对于研究细胞对机械生物学过程的反应所涉及的分子途径至关重要。准确评估这些反应需要对在这种细胞拉伸装置中培养的细胞进行详细观察。本研究旨在开发一种制备显微镜载玻片的方法，以便对这些装置中的细胞进行高倍成像。关键创新在于在细胞培养膜和上层通道之间形成可剥离的结合，从而便于去除上层并精确切割膜以进行高倍显微镜观察。使用制造的装置，对OP9细胞（每通道15,000个细胞）进行拉伸，并在配备100×和60×物镜的荧光显微镜下检查粘着斑蛋白的作用和YAP1的细胞内分布。拉伸刺激增加了OP9细胞中整合素β1的表达，并使整合素-纽蛋白复合物的形成增加了约1.4倍。此外，在拉伸过程中YAP1的核定位显著增强（约1.3倍）。该方法为使用基于微流控的细胞拉伸装置的研究人员提供了一种有价值的工具。预计微器件研究中成像技术的进步将进一步推动机械生物学研究的进展。
