Characterization of In Vitro Endothelial Linings Grown Within Microfluidic Channels
Characterization of In Vitro Endothelial Linings Grown Within Microfluidic Channels
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DOI：
10.1089/ten.tea.2010.0371
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发表时间：
2011-12-01
影响因子：
4.1
通讯作者：
Stokol, Tracy
中科院分区：
文献类型：
--
作者：
Esch, Mandy B.;Post, David J.;Stokol, Tracy
在体内，内皮细胞生长在血管内表面，其形状与血管的几何形状相符。在最小的血管中，这种形状意味着每个细胞内会有大幅弯曲。微加工通道能够复制这些小尺度的几何形状，但在其中生长的内皮细胞尚未得到充分的特性描述。特别是，在带有拐角的微通道中生长的内皮细胞中是否存在粘着斑和黏附连接尚未得到证实。我们在聚二甲基硅氧烷中制作了方形微流体通道（宽50微米，深50微米）和半圆形微流体通道（宽60微米，深45微米），并在其中培养人脐静脉内皮细胞（HUVEC）。通过共聚焦显微镜拍摄的图像堆栈的免疫荧光染色和三维重建证实，HUVEC能够在两种通道几何形状的所有通道壁上形成黏附连接，包括方形通道的侧壁。剪切应力的存在对于细胞在两种通道几何形状中形成粘着斑至关重要。剪切应力还促使HUVEC与通道壁贴合，并在方形通道内产生体外内皮衬里的方形横截面几何形状。因此，几何形状和施加的剪切应力是微血管体外内皮衬里开发的重要设计标准。
In vivo, endothelial cells grow on the inner surface of blood vessels and are shaped to conform to the vessel's geometry. In the smallest vessels this shape entails substantial bending within each cell. Microfabricated channels can replicate these small-scale geometries, but endothelial cells grown within them have not been fully characterized. In particular, the presence of focal adhesions and adherens junctions in endothelial cells grown in microchannels with corners has not been confirmed. We have fabricated square microfluidic channels (50 mu m wide, 50 mu m deep) and semicircular microfluidic channels (60 mu m wide, 45 mu m deep) in polydimethylsiloxane and cultured human umbilical vein endothelial cells (HUVEC) within them. Immunofluorescent staining and three-dimensional reconstruction of image stacks taken with confocal microscopy confirmed that HUVEC are capable of forming adherens junctions on all channel walls in both channel geometries, including the sidewalls of square profile channels. The presence of shear stress is critical for the cells to form focal adhesions within both channel geometries. Shear stress is also responsible for the conforming of HUVEC to the channel walls and produces a square cross-sectional geometry of in vitro endothelial linings within square profile channels. Thus, geometry and applied shear stress are important design criteria for the development of in vitro endothelial linings of microvessels.