Engineering high-density endothelial cell monolayers on soft substrates
Engineering high-density endothelial cell monolayers on soft substrates
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DOI：
10.1016/j.actbio.2009.01.032
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发表时间：
2009-07-01
影响因子：
9.7
通讯作者：
Brennan, Anthony B.
中科院分区：
文献类型：
--
作者：
Feinberg, Adam W.;Schumacher, James F.;Brennan, Anthony B.
这项研究表明，一个汇合单层的内皮细胞（EC）可以组织工程的软基板上的细胞密度和形态，接近体内条件。我们实现了在聚二甲基硅氧烷（PDMS）弹性体上的汇合EC单层的形成，纤连蛋白（FN）的微接触印刷在一个正方形点阵阵列的3 μ m直径的圆形岛屿在6 μ m的间距。FN或血清蛋白在PDMS或组织培养处理的聚苯乙烯上的均匀涂层未能支持等效EC密度和/或汇合。FN微图案化PDMS上的EC达到1，536 +/- 247个细胞mm（-2）的密度，接近于在猪肺动脉体内观察到的3，215 +/- 336个细胞mm（-2），并且显著高于（2- 5倍）其他材料上的EC密度。增强EC粘附、生长和密度的可能机制是增加EC与基底之间的粘着斑（FA）形成。培养14天后，微图案化的FN表面将每个细胞的FA平均数增加到35 +/-10，而均匀涂覆有FN的PDMS上的EC为7 +/-6。因此，FN在PDMS弹性体上形成FA大小的圆形岛的微尺度图案化促进了具有体内样细胞密度和形态的EC单层的形成。(C)2009 Acta Materialia Inc.由爱思唯尔有限公司出版。保留所有权利。
This study demonstrates that a confluent monolayer of endothelial cells (ECs) can be tissue engineered on a soft substrate with a cell density and morphology that approximates in vivo conditions. We achieved formation of a confluent EC monolayer on polydimethylsiloxane (PDMS) elastomer by microcontact printing of fibronectin (FN) in a square lattice array of 3 mu m diameter circular islands at a 6 mu m pitch. Uniform coatings of FN or serum proteins on PDMS or on tissue-culture-treated polystyrene failed to support the equivalent EC density and/or confluence. The ECs on the FN micropatterned PDMS achieved a density of 1,536 +/- 247 cells mm(-2), close to the 3,215 +/- 336 cells mm(-2) observed in vivo from porcine pulmonary artery and significantly higher (2- to 5-fold) than EC density on other materials. The probable mechanism for enhanced EC adhesion, growth and density is increased focal adhesion (FA) formation between the ECs and the substrate. After 14 days culture, the micropatterned FN surface increased the average number of FAs per cell to 35 +/- 10, compared to 7 +/- 6 for ECs on PDMS uniformly coated with FN. Thus, microscale patterning of FN into FA-sized, circular islands on PDMS elastomer promotes the formation of EC monolayers with in vivo-like cell density and morphology. (C) 2009 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.