Gradient patterned aECM proteins for cell studies

描述(申请人提供)：在体内，细胞整合了一组复杂的物理和化学信号以及这些信号的空间梯度，以维持适当的组织功能。在人工复制这些细胞微环境方面已经有了很大的进展，通过生产具有定制的表面化学成分或具有不同机械顺应性的区域的基质。然而，在允许对这两种性质进行独立调节以研究细胞反应的材料设计方面，进展甚微。我们建议通过制备具有叠加梯度的人工细胞外基质(AECM)薄膜并观察细胞在这些表面上的行为来应对这一挑战。这些独特的AECM蛋白由RGD和CS5的细胞结合域和弹性蛋白衍生的带有交联点的重复序列组成。化学线索中的梯度可以通过不同蛋白质的微流体混合来控制，而顺应性的梯度可以通过改变交联度来实现。这种特殊底物的制备为研究竞争信号如何产生协调的细胞反应提供了一种新的实验方法。这项工作的结果也可以应用于创造生物医学应用的改进材料。

DESCRIPTION (provided by applicant): In vivo, cells integrate a complex set of physical and chemical signals as well as spatial gradients of these signals to maintain proper tissue function. There has been much progress in the artificial replication of these cellular microenvironments by producing substrates with either tailored surface chemistries or regions of varying mechanical compliance. Yet there has been minimal advancement in the design of materials that allow for the independent modulation of both properties to study cellular response. We propose to address this challenge by fabricating artificial extracellular matrix (aECM) films with superimposed gradients and observing cellular behavior on these surfaces. These unique aECM proteins are comprised of cell binding domains of RGD and CS5 and an elastin-derived repeat with a crosslinking site. Gradients in chemical cues can be controlled by the microfluidic mixing of different proteins while gradients in compliance can be achieved by varying the extent of crosslinking. Preparation of this specialty substrate provides a novel experimental approach for examining how competing signals generate a coordinated cellular response. Results from this work can also be applied to create improved materials for biomedical applications.