Cell Biology Section, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland, USA.
Department of Biomedical Sciences, Elson S. Floyd College of Medicine, Washington State University, Spokane, Washington, USA.
Cancer Rep (Hoboken). 2020 Oct;3(5):e1257. doi: 10.1002/cnr2.1257. Epub 2020 Jun 8.
Three-dimensional (3D) in vitro model systems can bridge the gap between regular two-dimensional cell culture and whole-animal studies. Analyses of cancer cell migration and invasion increasingly use differing 3D systems, which may produce conflicting findings.
We directly compared different 3D extracellular matrix systems for studying cancer cell migration/invasion by analyzing cell morphologies and quantifying aspects of cell migration including speed and directional persistence using automated computer-based cell tracking.
We performed direct comparisons of five different 3D extracellular matrix cell culture systems using both HT1080 fibrosarcoma and MDA-MB-231 breast carcinoma cell lines. The reconstituted 3D systems included two types of collagen hydrogel and tissue matrix gel (TMG) vs cell-derived matrices extracted from cultured primary human or cancer-associated fibroblasts. The fibrillar matrix architecture of these systems differed. 3D rat tail collagen and TMG matrices had short, randomly oriented collagen fibrils; bovine collagen had long, larger fibril bundles; and the cell-derived matrices were strongly oriented. HT1080 cells displayed rounded morphologies in all three reconstituted 3D matrices but became spindle shaped in the two cell-derived matrices. MDA-MB-231 cell morphologies were elongated in all matrices. Quantitative measures of cell migration parameters differed markedly between the different types of 3D matrix. Comparing the reconstituted matrices, cells migrated the most rapidly and furthest in TMG. Comparing TMG with cell-derived matrices, cells migrated more efficiently in the cell-derived matrices. The most notable differences were in directional persistence of migration, which was greatest in the two cell-derived matrices.
The morphologies of matrix fibrils and cell shape, and particularly the efficiency and directionality of cell migration, differed substantially depending on the type of 3D matrix system. We suggest that it is important to employ the 3D model system that most closely resembles the matrix environment being studied for analyses of cancer cell migration and invasion.
三维（3D）体外模型系统可以弥合常规二维细胞培养与整体动物研究之间的差距。分析癌细胞的迁移和侵袭越来越多地使用不同的 3D 系统，这些系统可能会产生相互矛盾的结果。
我们通过分析细胞形态并使用自动计算机细胞跟踪量化细胞迁移的速度和方向持久性等方面，直接比较了不同的 3D 细胞外基质系统用于研究癌细胞迁移/侵袭的能力。
我们使用 HT1080 纤维肉瘤和 MDA-MB-231 乳腺癌细胞系，直接比较了五种不同的 3D 细胞外基质细胞培养系统。重建的 3D 系统包括两种类型的胶原水凝胶和组织基质凝胶（TMG）与从培养的原代人或癌症相关成纤维细胞中提取的细胞衍生基质。这些系统的纤维状基质结构不同。3D 大鼠尾胶原和 TMG 基质具有短而随机取向的胶原纤维；牛胶原具有长而更大的纤维束；而细胞衍生的基质则强烈取向。HT1080 细胞在所有三种重建的 3D 基质中均呈圆形形态，但在两种细胞衍生基质中则呈梭形。MDA-MB-231 细胞形态在所有基质中均呈细长形。细胞迁移参数的定量测量在不同类型的 3D 基质之间差异显著。比较重建的基质，细胞在 TMG 中迁移速度最快，距离最远。比较 TMG 与细胞衍生基质，细胞在细胞衍生基质中迁移效率更高。最显著的差异在于迁移的方向持久性，在两种细胞衍生基质中最大。
基质纤维的形态和细胞形状，特别是细胞迁移的效率和方向性，因 3D 基质系统的类型而异。我们建议，对于癌症细胞迁移和侵袭的分析，使用最接近所研究基质环境的 3D 模型系统非常重要。
