Division of Chemical Engineering, Graduate School of Engineering Science, Osaka University, Toyonaka, Osaka, Japan.
Biotechnol Lett. 2013 Jul;35(7):1001-8. doi: 10.1007/s10529-013-1174-x. Epub 2013 Mar 21.
Using a cell sheet stacking method, we developed an in vitro culture system in which green fluorescent protein expressing human umbilical vein endothelial cells (GFP-HUVECs) were cultured under human skeletal muscle myoblast (HSMM) sheets with different layer numbers. Our aim in developing this system was to examine the different endothelial behaviors in the cell sheet. During 96 h of incubation, in monolayer HSMM sheet, HUVECs quickly reached the top of the cell sheet and detached. In three-layered HSMM sheet, HUVECs also migrated to the top layer and formed island-shaped aggregates. In five-layered HSMM sheet, HUVECs migrated into the middle of the cell sheet and formed net-shaped aggregates. In seven-layered HSMM sheet, HUVECs migrated in the basal of the cell sheet and formed sparse net-shaped aggregates. The thickness of the HSMM sheet, which can be controlled by the layer number of the cell sheet, is therefore an important parameter that affects the migration time, encounters, localization, and morphology of HUVECs inside the HSMM sheet.
采用细胞片层堆叠方法，我们开发了一种体外培养体系，在该体系中，培养了表达绿色荧光蛋白的人脐静脉内皮细胞（GFP-HUVEC），并在具有不同层数的人骨骼肌成肌细胞（HSMM）片层下进行培养。我们开发该系统的目的是研究细胞片中不同的内皮细胞行为。在 96 小时的孵育过程中，在单层 HSMM 片层中，HUVEC 很快到达细胞片层的顶部并脱落。在三层 HSMM 片层中，HUVEC 也迁移到最上层并形成岛状聚集物。在五层 HSMM 片层中，HUVEC 迁移到细胞片层的中间并形成网状聚集物。在七层 HSMM 片层中，HUVEC 迁移到细胞片层的底部并形成稀疏的网状聚集物。HSMM 片层的厚度可以通过细胞片层的层数来控制，因此是影响 HUVEC 在内的迁移时间、相互作用、定位和形态的重要参数HSMM 片层。
