Storm Michael P, Sorrell Ian, Shipley Rebecca, Regan Sophie, Luetchford Kim A, Sathish Jean, Webb Steven, Ellis Marianne J
Department of Chemical Engineering and Centre for Regenerative Medicine, University of Bath;
MRC Centre for Drug Safety Science and Institute of Translational Medicine, University of Liverpool.
J Vis Exp. 2016 May 26(111):53431. doi: 10.3791/53431.
Tissue culture has been used for over 100 years to study cells and responses ex vivo. The convention of this technique is the growth of anchorage dependent cells on the 2-dimensional surface of tissue culture plastic. More recently, there is a growing body of data demonstrating more in vivo-like behaviors of cells grown in 3-dimensional culture systems. This manuscript describes in detail the set-up and operation of a hollow fiber bioreactor system for the in vivo-like culture of mammalian cells. The hollow fiber bioreactor system delivers media to the cells in a manner akin to the delivery of blood through the capillary networks in vivo. The system is designed to fit onto the shelf of a standard CO2 incubator and is simple enough to be set-up by any competent cell biologist with a good understanding of aseptic technique. The systems utility is demonstrated by culturing the hepatocarcinoma cell line HepG2/C3A for 7 days. Further to this and in line with other published reports on the functionality of cells grown in 3-dimensional culture systems the cells are shown to possess increased albumin production (an important hepatic function) when compared to standard 2-dimensional tissue culture.
组织培养已被用于体外研究细胞和反应超过100年。这项技术的传统做法是使贴壁依赖性细胞在组织培养塑料的二维表面上生长。最近,越来越多的数据表明,在三维培养系统中生长的细胞具有更类似体内的行为。本手稿详细描述了用于哺乳动物细胞类体内培养的中空纤维生物反应器系统的设置和操作。中空纤维生物反应器系统以类似于体内血液通过毛细血管网络输送的方式向细胞输送培养基。该系统设计为可安装在标准二氧化碳培养箱的架子上,简单到任何对无菌技术有良好理解的合格细胞生物学家都可以进行设置。通过培养肝癌细胞系HepG2/C3A 7天来证明该系统的实用性。除此之外,与其他关于在三维培养系统中生长的细胞功能的已发表报告一致,与标准二维组织培养相比,这些细胞显示出白蛋白产量增加(一项重要的肝功能)。