Tang Xiaoyun, Brindley David N
Department of Biochemistry, Cancer Research Institute of Northern Alberta, University of Alberta, Edmonton, AB, Canada.
Methods Mol Biol. 2022;2579:197-207. doi: 10.1007/978-1-0716-2736-5_15.
Three-dimensional (3D) cell culture technology is a powerful tool in cancer research and drug development. It retains several critical components of the in vivo environment, including cell-cell and cell-extracellular matrix (ECM) interactions in a 3D fashion, gradients of oxygen, nutrients and metabolic waste, and it is thus more physiologically relevant than traditional two-dimensional (2D) cell culture. Here, we describe a simple and versatile method using commercially available chamber slides and Matrigel, a surrogate ECM hydrogel, to set up a 3D culture model for breast cancer cells. In this 3D culture model, cells form aggregates or spheroids on top of a thin layer of Matrigel, which can be fixed directly onto the chamber slides for cell imaging and immunofluorescence staining. Alternatively, RNA and protein can be extracted from the cells for further investigation. This 3D cell culture model provides a useful platform for cancer research and drug development, in which the effects of novel compounds or genetic modifications can be tested.
三维（3D）细胞培养技术是癌症研究和药物开发中的一项强大工具。它保留了体内环境的几个关键组成部分，包括以三维方式进行的细胞-细胞和细胞-细胞外基质（ECM）相互作用、氧气、营养物质和代谢废物的梯度，因此比传统的二维（2D）细胞培养在生理上更具相关性。在此，我们描述一种简单且通用的方法，使用市售的培养室载玻片和基质胶（一种替代ECM水凝胶）来建立乳腺癌细胞的3D培养模型。在这个3D培养模型中，细胞在一层薄的基质胶上形成聚集体或球体，可直接固定在培养室载玻片上用于细胞成像和免疫荧光染色。或者，可以从细胞中提取RNA和蛋白质用于进一步研究。这种3D细胞培养模型为癌症研究和药物开发提供了一个有用的平台，可在其中测试新型化合物或基因修饰的效果。
