Department of Bioengineering, Speed School of Engineering, University of Louisville, Louisville, Kentucky 40208, USA.
Anal Chem. 2011 Apr 15;83(8):3170-7. doi: 10.1021/ac2002998. Epub 2011 Mar 17.
The phenotype and function of vascular cells in vivo are influenced by complex mechanical signals generated by pulsatile hemodynamic loading. Physiologically relevant in vitro studies of vascular cells therefore require realistic environments where in vivo mechanical loading conditions can be accurately reproduced. To accomplish a realistic in vivo-like loading environment, we designed and fabricated an Endothelial Cell Culture Model (ECCM) to generate physiological pressure, stretch, and shear stress profiles associated with normal and pathological cardiac flow states. Cells within this system were cultured on a stretchable, thin (∼500 μm) planar membrane within a rectangular flow channel and subject to constant fluid flow. Under pressure, the thin planar membrane assumed a concave shape, representing a segment of the blood vessel wall. Pulsatility was introduced using a programmable pneumatically controlled collapsible chamber. Human aortic endothelial cells (HAECs) were cultured within this system under normal conditions and compared to HAECs cultured under static and "flow only" (13 dyn/cm(2)) control conditions using microscopy. Cells cultured within the ECCM were larger than both controls and assumed an ellipsoidal shape. In contrast to static control control cells, ECCM-cultured cells exhibited alignment of cytoskeletal actin filaments and high and continuous expression levels of β-catenin indicating an in vivo-like phenotype. In conclusion, design, fabrication, testing, and validation of the ECCM for culture of ECs under realistic pressure, flow, strain, and shear loading seen in normal and pathological conditions was accomplished. The ECCM therefore is an enabling technology that allows for study of ECs under physiologically relevant biomechanical loading conditions in vitro.
体内血管细胞的表型和功能受脉动血流负荷产生的复杂机械信号的影响。因此，血管细胞的生理相关体外研究需要逼真的环境，能够准确再现体内机械加载条件。为了实现逼真的类似于体内的加载环境，我们设计并制造了一个内皮细胞培养模型（ECCM），以产生与正常和病理心脏血流状态相关的生理压力、拉伸和切应力分布。该系统中的细胞在可拉伸的、薄的（约 500μm）平面膜上培养，位于矩形流道内，并受到持续的流体流动。在压力下，薄的平面膜呈现出凹形，代表血管壁的一段。脉动是通过可编程气动控制可折叠室引入的。人主动脉内皮细胞（HAECs）在该系统中于正常条件下培养，并与静态和“仅流动”（13 dyn/cm2）对照条件下培养的 HAECs 通过显微镜进行比较。在 ECCM 中培养的细胞比对照细胞更大，并且呈椭圆形。与静态对照细胞不同，ECCM 培养的细胞表现出细胞骨架肌动蛋白丝的排列和β-连环蛋白的高且持续的表达水平，表明具有类似于体内的表型。总之，完成了用于在正常和病理条件下看到的真实压力、流动、应变和切变加载下培养 ECs 的 ECCM 的设计、制造、测试和验证。因此，ECCM 是一种使能技术，允许在体外生理相关生物力学加载条件下研究 ECs。
