Bakunts Karina, Gillum Nikki, Karabekian Zaruhi, Sarvazyan Narine
Pharmacology and Physiology Department, The George Washington University, Washington, DC 20037, USA.
Biotechniques. 2008 Mar;44(3):341-8. doi: 10.2144/000112682.
We report a simple in vitro model of cardiac tissue that mimics three-dimensional (3-D) environment and mechanical load conditions and, as such, may serve as a convenient method to study stem cell engraftment or address developmental questions such as cytoskeleton or intercalated disk maturation. To create in vitro cardiac fibers we used Matrigel, a commercially available basement membrane preparation. A semisolid pillow from concentrated Matrigel was overlaid with a suspension of rat neonatal cardiomyocytes in a diluted Matrigel solution. This created an environment in which the multicellular fibers continuously contracted against a mechanical load. The described approach allows continuous structural and functional monitoring of 20-300-micron-thick cardiac fibers and provides easy access to epitopes for immunostaining purposes.
我们报告了一种简单的心脏组织体外模型,该模型模拟三维(3-D)环境和机械负荷条件,因此可作为研究干细胞植入或解决诸如细胞骨架或闰盘成熟等发育问题的便捷方法。为了创建体外心脏纤维,我们使用了基质胶,一种市售的基底膜制剂。将浓缩基质胶制成的半固体枕状物覆盖在稀释的基质胶溶液中的大鼠新生心肌细胞悬液上。这创造了一个多细胞纤维在机械负荷下持续收缩的环境。所描述的方法允许对20-300微米厚的心脏纤维进行连续的结构和功能监测,并便于进行免疫染色以获取表位。