Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Ann Biomed Eng. 2011 Jun;39(6):1840-7. doi: 10.1007/s10439-011-0275-8. Epub 2011 Feb 19.
The differentiation process of murine embryonic stem cells into cardiomyocytes was investigated with a compliant microfluidic platform which allows for versatile cell seeding arrangements, optical observation access, long-term cell viability, and programmable uniaxial cyclic stretch. Specifically, two environmental cues were examined with this platform--culture dimensions and uniaxial cyclic stretch. First, the cardiomyogenic differentiation process, assessed by a GFP reporter driven by the α-MHC promoter, was enhanced in microfluidic devices (µFDs) compared with conventional well-plates. The addition of BMP-2 neutralizing antibody reduced the enhancement observed in the µFDs and the addition of exogenous BMP-2 augmented the cardiomyogenic differentiation in well plates. Second, 24 h of uniaxial cyclic stretch at 1 Hz and 10% strain on day 9 of differentiation was found to have a negative impact on cardiomyogenic differentiation. This microfluidic platform builds upon an existing design and extends its capability to test cellular responses to mechanical strain. It provides capabilities not found in other systems for studying differentiation, such as seeding embryoid bodies in 2D or 3D in combination with cyclic strain. This study demonstrates that the microfluidic system contributes to enhanced cardiomyogenic differentiation and may be a superior platform compared with conventional well plates. In addition to studying the effect of cyclic stretch on cardiomyogenic differentiation, this compliant platform can also be applied to investigate other biological mechanisms.
研究了一种顺应性微流控平台，该平台可用于各种细胞播种排列、光学观察通道、长期细胞活力和可编程的单轴循环拉伸，以研究鼠胚胎干细胞向心肌细胞的分化过程。具体来说，使用该平台研究了两个环境线索——培养尺寸和单轴循环拉伸。首先，通过由α-MHC 启动子驱动的 GFP 报告基因评估的心肌生成分化过程，在微流控装置（µFDs）中比传统的微孔板增强。在 µFDs 中添加 BMP-2 中和抗体减少了观察到的增强，而添加外源性 BMP-2 则增强了微孔板中的心肌生成分化。其次，在分化第 9 天以 1 Hz 和 10%应变施加 24 h 的单轴循环拉伸对心肌生成分化有负面影响。该微流控平台建立在现有设计的基础上，并扩展了其测试细胞对机械应变的响应能力。它提供了其他系统在研究分化时所没有的功能，例如在 2D 或 3D 中播种类胚体并结合循环应变。这项研究表明，微流控系统有助于增强心肌生成分化，并且与传统的微孔板相比可能是一个更好的平台。除了研究循环拉伸对心肌生成分化的影响外，这种顺应性平台还可用于研究其他生物学机制。
