Cell culture chips for simultaneous application of topographical and electrical cues enhance phenotype of cardiomyocytes
Cell culture chips for simultaneous application of topographical and electrical cues enhance phenotype of cardiomyocytes
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DOI：
10.1039/b810034a
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发表时间：
2009-01-01
期刊：
影响因子：
6.1
通讯作者：
Radisic, Milica
中科院分区：
文献类型：
--
作者：
Au, Hoi Ting Heidi;Cui, Bo;Radisic, Milica
在体内，心肌细胞暴露于多种生化和物理线索，包括地形和电线索。在标准组织培养装置中的长时间体外培养期间，已知心肌细胞由于缺乏适当的微环境线索而去分化。大多数目前可用的细胞培养系统仅提供单一的生物物理线索，因此迫切需要开发结合多种线索的先进细胞培养系统。我们在这里报告的发展，微加工系统，将地形和电线索在一个单一的芯片上，这使得分化的心肌细胞的培养。通过聚苯乙烯的热压印来产生细胞培养芯片，以产生精确限定的深度、宽度和周期性的微槽和微脊。研究了由0.5 μ m宽的槽和0.5 μ m宽的脊（1 μ m周期）组成的基底和由3 μ m宽的槽和1 μ m宽的脊（4 μ m周期）组成的基底，并将光滑表面用作对照。微槽的深度为400 nm。两个金电极电沉积1厘米，使微槽之间的取向平行或垂直于电极，使地形和电线索之间的相互作用的研究。新生大鼠心肌细胞上培养7天的微槽基板上延长和对齐沿着形成一个发达的收缩装置的微槽，证明了肌节α-辅肌动蛋白染色，与4 μ m相比，基板上的1 μ m的周期性更显着的效果。重要的是，同时应用双相电脉冲和地形线索导致间隙连接局限于细胞-细胞末端连接，而不是在新生儿细胞中发现的点状分布。当微槽平行于电场方向时，电场刺激进一步增强了心肌细胞的伸长。由于所描述的细胞培养芯片与荧光和光学显微镜的兼容性以及独立控制每个芯片上的场刺激参数、生物化学和地形学线索的能力，该系统将来可能成为药物开发和源自干细胞的心肌细胞成熟的有用工具。
In vivo, cardiomyocytes are exposed to multiple biochemical and physical cues including topographical and electrical cues. During prolonged in vitro cultivation in standard tissue culture set-ups, cardiomyocytes are known to de-differentiate due to the lack of appropriate micro-environmental cues. Most currently available cell culture systems provide only a single biophysical cue, thus development of advanced cell cultivation systems incorporating multiple cues is urgently needed. We report here the development of a microfabricated system, incorporating topographical and electrical cues on a single chip, which enables cultivation of differentiated cardiomyocytes. The cell culture chips were created by hot embossing of polystyrene, to create microgrooves and microridges of precisely defined depth, width and periodicity. Substrates consisting of 0.5 mu m-wide grooves and 0.5 mu m-wide ridges (1 mu m period) and those consisting of 3 mu m-wide grooves and 1 mu m-wide ridges (4 mu m period) were investigated, with smooth surfaces used as controls. The depth of the microgrooves was 400 nm. The two gold electrodes were electrodeposited 1 cm apart such that the microgrooves in-between were oriented either parallel or perpendicular to the electrodes, enabling studies of interaction between topographical and electrical cues. Neonatal rat cardiomyocytes cultivated on microgrooved substrates for 7 days were elongated and aligned along the microgrooves forming a well developed contractile apparatus, as evidenced by sarcomeric alpha-actinin staining, with a more pronounced effect on substrates with 1 mu m compared to 4 mu m periodicity. Importantly, simultaneous application of biphasic electrical pulses and topographical cues resulted in gap junctions confined to the cell-cell end junctions rather than the punctate distribution found in neonatal cells. Electrical field stimulation further enhanced cardiomyocyte elongation when microgrooves were oriented parallel to the electric field. Due to the compatibility of the described cell culture chips with fluorescence and optical microscopy as well as the ability to independently control field stimulation parameters, biochemical and topographical cues on each chip, this system may in the future become a useful tool in drug development and maturation of cardiomyocytes derived from stem cells.