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首页> 外文期刊>Cell communication & adhesion >Double-Layered Collagen Gel Hemisphere for Cell Invasion Assay: Successful Visualization and Quantification of Cell Invasion Activity
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Double-Layered Collagen Gel Hemisphere for Cell Invasion Assay: Successful Visualization and Quantification of Cell Invasion Activity

机译：双层胶原凝胶半球用于细胞侵袭测定：细胞侵袭活性的成功可视化和量化

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摘要

Although various methods for collagen gel-based cell invasion assays have been described, there continues to be a need for a simpler and more objective assay. Here, we describe an easy-to-prepare double-layered collagen gel hemisphere (DL-CGH) system that satisfies these requirements, and we demonstrate the advantages of this new system for visualizing cell movements during invasion. DL-CGH consists of a central core collagen layer surrounded by an outer cover collagen layer. A droplet of collagen I solution (containing cells to be examined) naturally forms a small hemisphere on the bottom of the culture dish. After this central core layer gels, a second droplet is placed atop the first gel, encapsulating it completely. The hemisphere is submerged in the medium and cultured. The invasive activity of cells that infiltrate from the inner to the outer layer can be evaluated optically. Using this in vitro system, we measured the inhibitory effect of E-cadherin expression on cancer cell invasion. DL-CGH also allowed visualization of interactions between invading cancer cells and the stroma. Cancer cells, which lack the proteases required for direct entrance into the three-dimensional collagen matrix, were seen to slip like amoebas through matrix gaps generated by the pericellular proteolytic activity of fibroblasts. [Supplementary materials are available for this article. Go to the publisher's online edition of Cell Communication and Adhesion for the following free supplemental resources: Movies 1-3; 4a and b].
机译：尽管已经描述了用于基于胶原凝胶的细胞侵袭测定的各种方法，但是仍然需要更简单和更客观的测定。在这里，我们描述了满足这些要求的易于制备的双层胶原凝胶半球（DL-CGH）系统，并且我们展示了这种新系统的优势，可用于可视化入侵过程中的细胞运动。 DL-CGH由被外层胶原蛋白层包围的中央核心胶原蛋白层组成。一滴胶原蛋白I溶液（包含待检查的细胞）自然在培养皿底部形成一个小半球。在该中央核心层胶凝之后，将第二滴液滴放置在第一胶凝剂的顶部，将其完全封装。将半球浸没在培养基中并进行培养。从内层到外层浸润的细胞的侵袭活性可以用光学方法评估。使用此体外系统，我们测量了E-钙粘蛋白表达对癌细胞侵袭的抑制作用。 DL-CGH还可以可视化入侵的癌细胞与基质之间的相互作用。癌细胞缺乏直接进入三维胶原蛋白基质所需的蛋白酶，人们发现它们像变形虫一样通过成纤维细胞的细胞周蛋白水解活性产生的基质间隙滑动。 [本文提供补充材料。有关以下免费补充资源，请访问发行商的在线版细胞通讯和粘附力在线版本：电影1-3;电影1-3。 4a和b]。

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