School of Engineering, Bernal Institute, University of Limerick, Limerick, Ireland.
School of Natural Sciences, Bernal Institute, University of Limerick, Limerick, Ireland; Health Research Institute, University of Limerick, Limerick, Ireland.
Biophys J. 2021 Mar 2;120(5):855-865. doi: 10.1016/j.bpj.2021.01.026. Epub 2021 Feb 3.
Microchannels are used as a transportation highway for suspended cells both in vivo and ex vivo. Lymphatic and cardiovascular systems transfer suspended cells through microchannels within the body, and microfluidic techniques such as lab-on-a-chip devices, flow cytometry, and CAR T-cell therapy utilize microchannels of similar sizes to analyze or separate suspended cells ex vivo. Understanding the forces that cells are subject to while traveling through these channels are important because certain applications exploit these cell properties for cell separation. This study investigated the influence that cytoskeletal impairment has on the inertial positions of circulating cells in laminar pipe flow. Two representative cancer cell lines were treated using cytochalasin D, and their inertial positions were investigated using particle streak imaging and compared between benign and metastatic cell lines. This resulted in a shift in inertial positions between benign and metastatic as well as treated and untreated cells. To determine and quantify the physical changes in the cells that resulted in this migration, staining and nanoindentation techniques were then used to determine the cells' size, circularity, and elastic modulus. It was found that the cells' exposure to cytochalasin D resulted in decreased elastic moduli of cells, with benign and metastatic cells showing decreases of 135 ± 91 and 130 ± 60 Pa, respectively, with no change in either size or shape. This caused benign, stiffer cancer cells to be more evenly distributed across the channel width than metastatic, deformable cancer cells; additionally, a decrease in the elastic moduli of both cell lines resulted in increased migration toward the channel center. These results indicate that the elastic modulus may play more of a part in the inertial migration of such cells than previously thought.
微通道被用作体内和体外悬浮细胞的运输高速公路。淋巴和心血管系统通过体内的微通道转移悬浮细胞,而微流控技术,如芯片上实验室设备、流式细胞术和 CAR T 细胞疗法,利用类似大小的微通道来分析或分离体外悬浮细胞。了解细胞在通过这些通道时所承受的力非常重要,因为某些应用利用这些细胞特性进行细胞分离。本研究调查了细胞骨架损伤对层流管道流动中循环细胞惯性位置的影响。使用细胞松弛素 D 处理两种代表性的癌细胞系,并使用粒子条纹成像技术研究它们的惯性位置,并在良性和转移性细胞系之间进行比较。这导致良性和转移性以及未处理和处理的细胞之间的惯性位置发生了转移。为了确定并量化导致这种迁移的细胞的物理变化,然后使用染色和纳米压痕技术来确定细胞的大小、圆度和弹性模量。结果发现,细胞暴露于细胞松弛素 D 导致细胞弹性模量降低,良性和转移性细胞的弹性模量分别降低了 135±91 和 130±60 Pa,而大小或形状均无变化。这使得良性、较硬的癌细胞比转移性、可变形的癌细胞更均匀地分布在通道宽度上;此外,两种细胞系的弹性模量降低都会导致向通道中心的迁移增加。这些结果表明,弹性模量在这些细胞的惯性迁移中可能比以前认为的更为重要。