Three-Dimensional Cell-Based Microarrays: Printing Pluripotent Stem Cells into 3D Microenvironments
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.
Key words3D cell microarray Pluripotent stem cells Cellular microenvironment High-content screening Cytotoxicity
The work involved in the development of this protocol was funded by NIH (ES020903) and NYSTEM (C026717). JFP acknowledges support from Fundação para a Ciência e Tecnologia (SFRH/BD/79590/2011), which also funded iBB (UID/BIO/04565/2013).