Micropatterned cell cultures on elastic membranes as an in vitro model of myocardium
Micropatterned cell cultures on elastic membranes as an in vitro model of myocardium
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DOI：
10.1038/nprot.2006.203
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发表时间：
2006-01-01
期刊：
影响因子：
14.8
通讯作者：
McCulloch, Andrew D.
中科院分区：
文献类型：
--
作者：
Camelliti, Patrizia;Gallagher, John O.;McCulloch, Andrew D.
我们在这里描述了一种新的体外方案，通过控制培养细胞的位置和机械环境来模拟体内心室心肌表型的重要方面。微光刻技术用于制造微结构金属硅晶圆。这些用于制造微槽硅胶膜或硅胶模具，用于细胞外基质蛋白在弹性膜上的微流体应用（涉及微米分辨率的流量控制）。物理或微流体结构膜作为细胞培养生长基质，支持细胞排列并允许拉伸应用。后者是通过可提供各向同性或各向异性拉伸的拉伸装置实现的。在这些微图案膜上生长的新生儿心室心肌细胞，形成具有规则肌瘤图案的活体样形态。从制造微图纹金属硅晶圆到铸造硅胶模具、微流体图纹和细胞分离和播种的整个过程大约需要7天。
We describe here a new in vitro protocol for structuring cardiac cell cultures to mimic important aspects of the in vivo ventricular myocardial phenotype by controlling the location and mechanical environment of cultured cells. Microlithography is used to engineer microstructured silicon metal wafers. Those are used to fabricate either microgrooved silicone membranes or silicone molds for microfluidic application of extracellular matrix proteins onto elastic membranes (involving flow control at micrometer resolution). The physically or microfluidically structured membranes serve as a cell culture growth substrate that supports cell alignment and allows the application of stretch. The latter is achieved with a stretching device that can deliver isotropic or anisotropic stretch. Neonatal ventricular cardiomyocytes, grown on these micropatterned membranes, develop an in vivo-like morphology with regular sarcomeric patterns. The entire process from fabrication of the micropatterned silicon metal wafers to casting of silicone molds, microfluidic patterning and cell isolation and seeding takes approximately 7 days.