McLane Joshua S, Schaub Nicholas J, Gilbert Ryan J, Ligon Lee A
Department of Biology, Rensselaer Polytechnic Institute, Troy, NY, USA.
Methods Mol Biol. 2013;1046:371-88. doi: 10.1007/978-1-62703-538-5_23.
It has become increasingly clear that the cellular microenvironment, in particular the extracellular matrix, plays an important role in regulating cell function. However, the extracellular matrix is extraordinarily complex in both its makeup and its physical properties. Therefore, there is a need to develop model systems to independently evaluate the effect of specific extracellular matrix features upon cells. Here we describe a model system to evaluate one aspect of the extracellular matrix, its fibrous topology. We describe how to generate bio-mimetic nanofibers by electrospinning, how to grow cells on these fibers, and also some methods for fixing and visualizing cells grown on these fibers. These methods can be used to investigate a wide range of biological questions, including, but not limited to, cell-extracellular matrix adhesion and cell motility on extracellular matrix.
越来越明显的是,细胞微环境,尤其是细胞外基质,在调节细胞功能方面起着重要作用。然而,细胞外基质在其组成和物理性质方面都极其复杂。因此,需要开发模型系统来独立评估特定细胞外基质特征对细胞的影响。在这里,我们描述了一个模型系统,用于评估细胞外基质的一个方面,即其纤维拓扑结构。我们描述了如何通过静电纺丝生成仿生纳米纤维,如何在这些纤维上培养细胞,以及一些固定和可视化在这些纤维上生长的细胞的方法。这些方法可用于研究广泛的生物学问题,包括但不限于细胞与细胞外基质的粘附以及细胞在细胞外基质上的运动。