A matrix micropatterning platform for cell localization and stem cell fate determination.
A matrix micropatterning platform for cell localization and stem cell fate determination.
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DOI：
10.1016/j.actbio.2010.06.033
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发表时间：
2010-12
影响因子：
9.7
通讯作者：
Cooke, John P.
中科院分区：
文献类型：
--
作者：
Huang, Ngan F.;Patlolla, Bhagat;Abilez, Oscar;Sharma, Himanshu;Rajadas, Jaykumar;Beygui, Ramin E.;Zarins, Christopher K.;Cooke, John P.
为了研究细胞-ECM相互作用的作用，微尺度方法提供了对ECM微环境对细胞功能和表型进行高通量评估的潜力。使用微尺度直写（MDW）技术，我们的特点是多组分ECM微阵列的细胞微图案化，定位和干细胞命运的决定。将各种几何形状和尺寸的ECM和其他生物分子印刷到环氧化物改性的玻璃基底上，用于评价人内皮细胞的细胞附着。内皮细胞表现出强烈的优先附着ECM图案的区域，并对齐他们的细胞骨架沿着方向的微图案。我们接下来生成了含有一种或多种ECM组合物（即明胶、胶原IV和纤连蛋白）的ECM微阵列，然后在微阵列上培养鼠胚胎干细胞（ESC）。当维持在含有白血病抑制因子的生长培养基中时，ESC选择性地附着于微图案特征并表达与多能表型相关的标记物，例如E-钙粘蛋白和碱性磷酸酶。在可溶性因子视黄酸和骨形态发生蛋白-4的存在下，ESCs在ECM芯片上向外胚层谱系分化，具有差异ECM效应。在明胶上培养的ESC显示出显着更高水平的泛细胞角蛋白的表达，当比较培养的细胞胶原IV或纤连蛋白，这表明明胶优先促进外胚层分化。总之，我们的研究结果表明，MDW是一种通用的方法，打印不同的几何形状和组成的ECM到表面上，它是适合的多组分ECM微阵列的生成干细胞命运的决定。
To study the role of cell–ECM interactions, microscale approaches provide the potential to perform high throughput assessment of ECM microenvironments on cellular function and phenotype. Using a microscale direct writing (MDW) technique, we characterized the generation of multicomponent ECM microarrays for cellular micropatterning, localization, and stem cell fate determination. ECMs and other biomolecules of various geometries and sizes were printed onto epoxide-modified glass substrates for evaluation of cell attachment by human endothelial cells. The endothelial cells displayed strong preferential attachment to the ECM-patterned regions and aligned their cytoskeleton along the direction of the micropatterns. We next generated ECM microarrays that contained one or more ECM compositions (namely gelatin, collagen IV, and fibronectin) and then cultured murine embryonic stem cell (ESCs) on the microarrays. The ESCs selectively attached to the micropatterned features and expressed markers associated with a pluripotent phenotype, such as E-cadherin and alkaline phosphatase, when maintained in growth media containing leukemia inhibitory factor. In the presence of soluble factors retinoic acid and bone morphogenetic protein-4, the ESCs differentiated towards ectodermal lineage on the ECM microarray with differential ECM effects. The ESCs cultured on gelatin showed significantly higher levels of pan cytokeratin expression, when compared cells cultured on collagen IV or fibronectin, suggesting that gelatin preferentially promotes ectodermal differentiation. In summary, our results demonstrate that MDW is a versatile approach to print ECMs of diverse geometries and compositions onto surfaces, and it is amenable to the generation of multicomponent ECM microarrays for stem cell fate determination.
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DOI：
10.1039/b815718a
发表时间：
2009-01
期刊：
Integrative biology : quantitative biosciences from nano to macro
影响因子：
--
作者：
Lutolf MP;Doyonnas R;Havenstrite K;Koleckar K;Blau HM
通讯作者：
Blau HM
影响因子：
5.2
作者：
Hayashi, Yohei;Furue, Miho Kusuda;Asashima, Makoto
通讯作者：
Asashima, Makoto
影响因子：
56.9
作者：
Piner, RD;Zhu, J;Mirkin, CA
通讯作者：
Mirkin, CA
影响因子：
13.3
作者：
Mei, Ying;Cannizzaro, Christopher;Anderson, Daniel G.
通讯作者：
Anderson, Daniel G.
影响因子：
14
作者：
Roth, EA;Xu, T;Boland, T
通讯作者：
Boland, T