Stiffness of the substrate influences the phenotype of embryonic chicken cardiac myocytes
Stiffness of the substrate influences the phenotype of embryonic chicken cardiac myocytes
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DOI：
10.1002/jbm.a.32951
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发表时间：
2010-12-15
影响因子：
4.9
通讯作者：
Bashir, Rashid
中科院分区：
文献类型：
--
作者：
Bajaj, Piyush;Tang, Xin;Bashir, Rashid
我们研究了基板刚度的搏动率，收缩力和细胞骨架结构的胚胎鸡心肌细胞培养层粘连蛋白包被的聚丙烯酰胺（PA）基板上的效果。PA基板上培养的细胞与弹性的天然心肌（18千帕）表现出最高的跳动率在培养的头几天。单个细胞在所有基质上的初始搏动速率变化显著，但在5天内开始收敛。我们还研究了粘着斑（FA）和细胞骨架结构上不同的基板，通过共聚焦显微镜，并发现了较高的百分比FA组织培养（TC）塑料皿相比，较软的PA凝胶。此外，在18 kPa、50 kPa和TC培养皿上，高度排列的肌节条纹清晰可见，而在1 kPa培养皿上，细胞仅显示不排列的弥散条纹。使用微机电系统力传感器测量这些基底上的收缩力，其显示TC培养皿上的细胞的收缩力（F = 71.30 +/- 6.38 nN）显著大于在18-kPa PA凝胶上培养的细胞的收缩力（F = 30.16 +/- 3.83 nN）。这很可能是由于TC培养皿上形成的FA百分比较高，而较软凝胶上形成的FA较少。我们的累积研究结果可能对三维心脏组织工程支架的设计产生重大影响。(C)2010 Wiley Periodicals，Inc. J Biomed Mater Res Part A：95A：1261- 1269，2010.
We examined the effect of substrate stiffness on the beating rate, force of contraction, and cytoskeletal structure of embryonic chicken cardiac myocytes by culturing them on laminin-coated polyacrylamide (PA) substrates. Cells cultured on PA substrates with elasticity comparable to that of the native myocardium (18 kPa) exhibited the highest beating rate during the first few days of culture. The initial beating rate of individual cells on all the substrates varied significantly but began to converge within 5 days. We also examined the focal adhesions (FAs) and cytoskeletal structure on different substrates via confocal microscopy and found a higher percentage of FAs on tissue culture (TC) plastic dishes compared with the softer PA gels. Furthermore, highly aligned sarcomeric striations were clearly visible on 18 kPa, 50 kPa, and TC dish, whereas cells on 1 kPa only showed nonaligned diffused striations. The force of contraction on these substrates was measured using a micro-electromechanical system force sensor, which showed that the force of contraction for the cells on TC dishes (F = 71.30 +/- 6.38 nN) was significantly larger than those cultured on the 18-kPa PA gel (F = 30.16 +/- 3.83 nN). This is most likely due to the formation of higher percentage of FAs on the TC dishes compared with fewer FAs on the softer gels. Our cumulative findings can have a significant impact on the design of 3D cardiac tissue engineered scaffolds. (C) 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 95A: 1261-1269,2010.