Nested collagen matrices: A new model to study migration of human fibroblast populations in three dimensions
Nested collagen matrices: A new model to study migration of human fibroblast populations in three dimensions
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DOI：
10.1016/j.yexcr.2005.010.001
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发表时间：
2006-01-01
影响因子：
3.7
通讯作者：
Jiang, HM
中科院分区：
文献类型：
--
作者：
Grinnell, F;Rocha, LB;Jiang, HM
成纤维细胞-3D胶原基质培养提供了一种模型系统，用于在比常规2D细胞培养更接近组织的条件下分析细胞生理学。以前的工作主要集中在成纤维细胞对胶原基质的重塑和收缩，而对基质内细胞群迁移的研究很少。在这里，我们介绍了一个嵌套的胶原蛋白基质模型来分析成纤维细胞在3D胶原蛋白基质中的迁移。通过将收缩的含细胞基质（也称为真皮等同物）包埋在无细胞基质内制备嵌套胶原基质;发生从前者到后者的迁移。用人类皮肤碎片代替皮肤等同物的对照实验证实了该模型的可靠性。人成纤维细胞迁移在嵌套的胶原蛋白基质中发生在8-16小时的滞后期后，并且迁移出内部基质的细胞是双极的，具有领先的树突状延伸。迁移是肌球蛋白11，Rho激酶和金属蛋白酶依赖，但不需要血浆纤连蛋白。血小板衍生生长因子而不是溶血磷脂酸或血清刺激细胞迁移，尽管所有这三种生理激动剂促进基质重塑和收缩。巢式胶原基质模型是一种相对简单、快速和定量测量细胞群迁移的方法。我们使用该模型的研究证明了成纤维细胞迁移和胶原基质重塑的调节之间的重要差异。(c)2005年爱思唯尔公司All rights reserved.
Fibroblast-3D collagen matrix culture provides a model system to analyze cell physiology under conditions that more closely resemble tissue than conventional 2D cell culture. Previous work has focused primarily on remodeling and contraction of collagen matrices by fibroblasts, and there has been little research on migration of cell populations within the matrix. Here, we introduce a nested collagen matrix model to analyze migration of fibroblasts in 3D collagen matrices. Nested collagen matrices were prepared by embedding contracted cell-containing matrices (also called dermal equivalents) inside cell-free matrices; migration occurred from the former to the latter. Control experiments with human dermal fragments in place of dermal equivalents confirmed the reliability of the model. Human fibroblast migration in nested collagen matrices occurred after a lag phase of 8-16 h, and cells migrating out of the inner matrices were bipolar with leading dendritic extensions. Migration was myosin 11, Rho kinase and metalloproteinase-dependent but did not require plasma fibronectin. Platelet-derived growth factor but not lysophosphatidic acid or serum stimulated cell migration, although all three of these physiological agonists promote matrix remodeling and contraction. The nested collagen matrix model is a relatively easy, rapid and quantitative method to measure migration of cell populations. Our studies using this model demonstrate important differences between regulation of fibroblast migration and remodeling in collagen matrices. (c) 2005 Elsevier Inc. All rights reserved.