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利用基因工程改造的、与金和细胞结合的多肽创建细胞模式。

Creating cellular patterns using genetically engineered, gold- and cell-binding polypeptides.

作者信息

Li Linying, Mo Chia-Kuei, Chilkoti Ashutosh, Lopez Gabriel P, Carroll Nick J

机构信息

Department of Biomedical Engineering, Duke University, Durham, North Carolina 27708 and Research Triangle Materials Research Science and Engineering Center (MRSEC), Durham, North Carolina 27708.

Department of Biomedical Engineering, Duke University, Durham, North Carolina 27708.

出版信息

Biointerphases. 2016 Jun 27;11(2):021009. doi: 10.1116/1.4952452.

Abstract

Patterning cells on material surfaces is an important tool for the study of fundamental cell biology, tissue engineering, and cell-based bioassays. Here, the authors report a simple approach to pattern cells on gold patterned silicon substrates with high precision, fidelity, and stability. Cell patterning is achieved by exploiting adsorbed biopolymer orientation to either enhance (gold regions) or impede (silicon oxide regions) cell adhesion at particular locations on the patterned surface. Genetic incorporation of gold binding domains enables C-terminal chemisorption of polypeptides onto gold regions with enhanced accessibility of N-terminal cell binding domains. In contrast, the orientation of polypeptides adsorbed on the silicon oxide regions limit the accessibility of the cell binding domains. The dissimilar accessibility of cell binding domains on the gold and silicon oxide regions directs the cell adhesion in a spatially controlled manner in serum-free medium, leading to the formation of well-defined cellular patterns. The cells are confined within the polypeptide-modified gold regions and are viable for eight weeks, suggesting that bioactive polypeptide modified surfaces are suitable for long-term maintenance of patterned cells. This study demonstrates an innovative surface-engineering approach for cell patterning by exploiting distinct ligand accessibility on heterogeneous surfaces.

摘要

在材料表面对细胞进行图案化是研究基础细胞生物学、组织工程和基于细胞的生物测定的重要工具。在此,作者报告了一种在金图案化硅基板上高精度、高保真度和高稳定性地对细胞进行图案化的简单方法。细胞图案化是通过利用吸附的生物聚合物取向来实现的,即在图案化表面的特定位置增强(金区域)或阻碍(氧化硅区域)细胞粘附。金结合域的基因掺入使得多肽能够通过C端化学吸附到金区域,同时增强N端细胞结合域的可及性。相比之下,吸附在氧化硅区域的多肽的取向限制了细胞结合域的可及性。金和氧化硅区域上细胞结合域的不同可及性在无血清培养基中以空间控制的方式引导细胞粘附,从而导致形成明确的细胞图案。细胞被限制在多肽修饰的金区域内,并且可以存活八周,这表明生物活性多肽修饰的表面适用于图案化细胞的长期维持。这项研究展示了一种创新的表面工程方法,通过利用异质表面上不同的配体可及性来进行细胞图案化。

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