Department of Bioengineering, Speed School of Engineering, University of Louisville, Louisville, Kentucky 40208, USA.
Anal Chem. 2010 Sep 15;82(18):7581-7. doi: 10.1021/ac1012893.
Physiological heart development and cardiac function rely on the response of cardiac cells to mechanical stress during hemodynamic loading and unloading. These stresses, especially if sustained, can induce changes in cell structure, contractile function, and gene expression. Current cell culture techniques commonly fail to adequately replicate physical loading observed in the native heart. Therefore, there is a need for physiologically relevant in vitro models that recreate mechanical loading conditions seen in both normal and pathological conditions. To fulfill this need, we have developed a microfluidic cardiac cell culture model (μCCCM) that for the first time allows in vitro hemodynamic stimulation of cardiomyocytes by directly coupling cell structure and function with fluid induced loading. Cells are cultured in a small (1 cm diameter) cell culture chamber on a thin flexible silicone membrane. Integrating the cell culture chamber with a pump, collapsible pulsatile valve and an adjustable resistance element (hemostatic valve) in series allow replication of various loading conditions experienced in the heart. This paper details the design, modeling, fabrication and characterization of fluid flow, pressure and stretch generated at various frequencies to mimic hemodynamic conditions associated with the normal and failing heart. Proof-of-concept studies demonstrate successful culture of an embryonic cardiomyoblast line (H9c2 cells) and establishment of an in vivo like phenotype within this system.
生理心脏发育和心功能依赖于心脏细胞在血液动力学加载和卸载过程中对机械应激的反应。这些应激，特别是如果持续存在，会引起细胞结构、收缩功能和基因表达的变化。目前的细胞培养技术通常无法充分复制天然心脏中观察到的物理加载。因此，需要生理相关的体外模型来重现正常和病理条件下观察到的机械加载条件。为了满足这一需求，我们开发了一种微流控心脏细胞培养模型（μCCCM），该模型首次通过直接将细胞结构和功能与流体诱导的加载相耦合，实现了对心肌细胞的体外血液动力学刺激。细胞在一个小的（1 厘米直径）细胞培养室内培养在一个薄的柔性硅树脂膜上。将细胞培养室与泵、可折叠脉动阀和可调电阻元件（止血阀）串联集成，允许复制心脏中经历的各种加载条件。本文详细介绍了在各种频率下产生的流体流动、压力和拉伸的设计、建模、制造和特性，以模拟与正常和衰竭心脏相关的血液动力学条件。概念验证研究表明，该系统成功培养了一种胚胎心肌细胞系（H9c2 细胞），并建立了类似于体内的表型。
