Pascoal Jorge F, Fernandes Tiago G, Nierode Gregory J, Diogo Maria Margarida, Dordick Jonathan S, Cabral Joaquim M S
Department of Bioengineering, Institute for Bioengineering and Biosciences (iBB), Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal.
Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, USA.
Methods Mol Biol. 2018;1771:69-81. doi: 10.1007/978-1-4939-7792-5_6.
Cell-based microarrays are valuable platforms for the study of cytotoxicity and cellular microenvironment because they enable high-throughput screening of large sets of conditions at reduced reagent consumption. However, most of the described microarray technologies have been applied to two-dimensional cultures, which do not accurately emulate the in vivo three-dimensional (3D) cell-cell and cell-extracellular matrix interactions.Herein, we describe the methodology for production of alginate- and Matrigel-based 3-D cell microarrays for the study of mouse and human pluripotent stem cells on two different chip-based platforms. We further provide protocols for on-chip proliferation/viability analysis and the assessment of protein expression by immunofluorescence.
基于细胞的微阵列是研究细胞毒性和细胞微环境的重要平台，因为它们能够在减少试剂消耗的情况下对大量条件进行高通量筛选。然而，大多数已描述的微阵列技术已应用于二维培养，而二维培养不能准确模拟体内三维（3D）细胞-细胞和细胞-细胞外基质相互作用。在此，我们描述了在两种不同的基于芯片的平台上生产用于研究小鼠和人类多能干细胞的基于藻酸盐和基质胶的3D细胞微阵列的方法。我们还提供了芯片上增殖/活力分析以及通过免疫荧光评估蛋白质表达的方案。
