Three-dimensional photolithographic micropatterning: a novel tool to probe the complexities of cell migration.
Three-dimensional photolithographic micropatterning: a novel tool to probe the complexities of cell migration.
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DOI：
10.1039/c3ib20280a
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发表时间：
2013-05
期刊：
影响因子：
--
通讯作者：
West JL
中科院分区：
文献类型：
--
作者：
Hoffmann JC;West JL
为了独立研究影响细胞运动的众多变量，有必要采用新颖的工具和材料来精细控制细胞微环境。在这项工作中，我们将先进的3D微图图化技术，即双光子激光扫描光刻技术（TP-LSL）应用于生物活性肽修饰的聚乙二醇（PEG）水凝胶，以制造精确设计的微环境，以指导和定量研究细胞迁移。具体而言，TP-LSL用于在不可降解的peg基水凝胶表面（2D）和可水解的peg基水凝胶内部（3D）制备细胞粘附PEG-RGDS微图案。通过延时显微镜观察，HT1080细胞在二维和三维情况下沿着这些粘附微模式迁移。根据粘附配体、水凝胶组成和二维和三维迁移的图案面积的变化，观察到细胞速度、细胞持久性和细胞形状的差异。结果表明，HT1080s在二维表面上的迁移速度更快，持久性较低，而在三维表面上迁移的HT1080s更小，更长。此外，细胞迁移显示出对PEG-RGDS浓度的双相依赖性，并且随着时间的推移，在PEG-RGDS微模式内移动的细胞移动得更快，更持久。重要的是，这里提出的工作开始阐明涉及细胞迁移的多个复杂因素，典型的混杂因素是独立控制的。这种独特平台的开发将使研究人员能够探索细胞在日益复杂的3D微环境中的行为，这些微环境开始模仿体内景观的特定选择方面。
In order to independently study the numerous variables that influence cell movement, it will be necessary to employ novel tools and materials that allow for exquisite control of the cellular microenvirenment. In this work, we have applied advanced 3D micropatterning technology, known as two-photon laser scanning lithography (TP-LSL), to poly(ethylene glycol) (PEG) hydrogels modified with bioactive peptides in order to fabricate precisely designed microenvirenments to guide and quantitatively investigate cell migration. Specifically, TP-LSL was used to fabricate cell adhesive PEG-RGDS micropatterns on the surface of non-degradable PEG-based hydrogels (2D) and in the interior of proteolytically degradable PEG-based hydrogels (3D). HT1080 cell migration was guided down these adhesive micropatterns in both 2D and 3D, as observed via time-lapse microscopy. Differences in cell speed, cell persistence, and cell shape were observed based on variation of adhesive ligand, hydrogel composition, and patterned area for both 2D and 3D migration. Results indicated that HT1080s migrate faster and with lower persistence on 2D surfaces, while HT1080s migrating in 3D were smaller and more elongated. Further, cell migration was shown to have a biphasic dependence on PEG-RGDS concentration and cells moving within PEG-RGDS micropatterns were seen to move faster and with more persistence over time. Importantly, the work presented here begins to elucidate the multiple complex factors involved in cell migration, with typical confounding factors being independently controlled. The development of this unique platform will allow researchers to probe how cells behave within increasingly complex 3D microenvironments that begin to mimic specifically chosen aspects of the in vivo landscape.
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