Timpson Paul, McGhee Ewan J, Erami Zahra, Nobis Max, Quinn Jean A, Edward Mike, Anderson Kurt I
The Beatson Institute for Cancer Research, University of Glasgow.
J Vis Exp. 2011 Oct 13(56):e3089. doi: 10.3791/3089.
Cell migration is fundamental to many aspects of biology, including development, wound healing, the cellular responses of the immune system, and metastasis of tumor cells. Migration has been studied on glass coverslips in order to make cellular dynamics amenable to investigation by light microscopy. However, it has become clear that many aspects of cell migration depend on features of the local environment including its elasticity, protein composition, and pore size, which are not faithfully represented by rigid two dimensional substrates such as glass and plastic. Furthermore, interaction with other cell types, including stromal fibroblasts and immune cells, has been shown to play a critical role in promoting the invasion of cancer cells. Investigation at the molecular level has increasingly shown that molecular dynamics, including response to drug treatment, of identical cells are significantly different when compared in vitro and in vivo. Ideally, it would be best to study cell migration in its naturally occurring context in living organisms, however this is not always possible. Intermediate tissue culture systems, such as cell derived matrix, matrigel, organotypic culture (described here) tissue explants, organoids, and xenografts, are therefore important experimental intermediates. These systems approximate certain aspects of an in vivo environment but are more amenable to experimental manipulation such as use of stably transfected cell lines, drug treatment regimes, long term and high-resolution imaging. Such intermediate systems are especially useful as proving grounds to validate probes and establish parameters required to image the dynamic response of cells and fluorescent reporters prior to undertaking imaging in vivo. As such, they can serve an important role in reducing the need for experiments on living animals.
细胞迁移是生物学许多方面的基础，包括发育、伤口愈合、免疫系统的细胞反应以及肿瘤细胞的转移。为了使细胞动力学便于通过光学显微镜进行研究，人们在玻璃盖玻片上对迁移进行了研究。然而，很明显，细胞迁移的许多方面取决于局部环境的特征，包括其弹性、蛋白质组成和孔径，而玻璃和塑料等刚性二维基质并不能如实地体现这些特征。此外，与其他细胞类型（包括基质成纤维细胞和免疫细胞）的相互作用已被证明在促进癌细胞侵袭方面起着关键作用。分子水平的研究越来越表明，相同细胞在体外和体内的分子动力学（包括对药物治疗的反应）存在显著差异。理想情况下，最好在生物体的自然环境中研究细胞迁移，但这并不总是可行的。因此，中间组织培养系统，如细胞衍生基质、基质胶、器官型培养（本文所述）、组织外植体、类器官和异种移植，是重要的实验中间体。这些系统近似于体内环境的某些方面，但更便于进行实验操作，如使用稳定转染的细胞系、药物治疗方案、长期和高分辨率成像。这种中间系统作为验证探针和确定在体内成像之前对细胞和荧光报告分子的动态反应进行成像所需参数的试验场特别有用。因此，它们在减少对活体动物实验的需求方面可以发挥重要作用。
