Department of Mechanical Engineering, National Taiwan University of Science and Technology, Taipei 106, Taiwan.
Biosensors (Basel). 2022 Mar 27;12(4):199. doi: 10.3390/bios12040199.
The cell membrane permeability of a cell type to water (Lp) and cryoprotective agents (Ps), is the key factor that determines the optimal cooling and mass transportation during cryopreservation. The human lung adenocarcinoma cell line, CL1, has been widely used to study the invasive capabilities or drug resistance of lung cancer cells. Therefore, providing accurate databases of the mass transport properties of this specific cell line can be crucial for facilitating either flexible and optimal preservation, or supply. In this study, utilizing our previously proposed noncontact-based micro-vortex system, we focused on comparing the permeability phenomenon between CL1-0 and its more invasive subline, CL1-5, under several different ambient temperatures. Through the assay procedure, the cells of favor were virtually trapped in a hydrodynamic circulation to provide direct inspection using a high-speed camera, and the images were then processed to achieve the observation of a cell's volume change with respect to time, and in turn, the permeability. Based on the noncontact nature of our system, we were able to manifest more accurate results than their contact-based counterparts, excluding errors involved in estimating the cell geometry. As the results in this experiment showed, the transport phenomena in the CL1-0 and CL1-5 cell lines are mainly composed of simple diffusion through the lipid bilayer, except for the case where CL1-5 were suspended in the cryoprotective agent (CPA) solution, which also demonstrated higher Ps values. The deviated behavior of CL1-5 might be a consequence of the altered expression of aquaporins and the coupling of a cryoprotective agent and water, and has given a vision on possible studies over these properties, and their potential relationship to invasiveness and metastatic stability of the CL1 cell line.
细胞膜对水（Lp）和冷冻保护剂（Ps）的通透性是决定冷冻保存过程中最佳冷却和质量传输的关键因素。人肺腺癌细胞系 CL1 已广泛用于研究肺癌细胞的侵袭能力或耐药性。因此，提供该特定细胞系质量传输特性的准确数据库对于促进灵活和最佳的保存或供应可能至关重要。在这项研究中，我们利用先前提出的非接触式微涡系统，重点比较了在几种不同环境温度下，CL1-0 和其侵袭性更强的亚系 CL1-5 之间的通透性现象。通过实验程序，有利的细胞实际上被困在流体动力学循环中，使用高速摄像机进行直接检查，然后处理图像以实现观察细胞体积随时间的变化，从而实现渗透率的观察。基于我们系统的非接触性质，我们能够获得比接触式对应物更准确的结果，排除了在估计细胞几何形状时涉及的误差。如实验结果所示，CL1-0 和 CL1-5 细胞系中的传输现象主要由穿过脂质双层的简单扩散组成，除了 CL1-5 悬浮在冷冻保护剂（CPA）溶液中的情况外，这种情况还显示出更高的 Ps 值。CL1-5 的偏离行为可能是水通道蛋白表达改变和冷冻保护剂与水偶联的结果，并为研究这些特性及其与 CL1 细胞系侵袭性和转移性稳定性的潜在关系提供了一个视角。
