%0 Journal Article %J Nat Protoc %D 2020 %T Activation and expansion of human T cells using artificial antigen-presenting cell scaffolds %A Zhang, David K Y %A Cheung, Alexander S %A Mooney, David J %K Antigen-Presenting Cells %K Cell Lineage %K Cloning, Molecular %K Gene Expression Regulation %K Humans %K T-Lymphocytes %X Synthetic antigen-presenting cells (APCs) are used to mediate scalable ex vivo T-cell expansion for adoptive cell therapy. Recently, we developed APC-mimetic scaffolds (APC-ms), which present signals to T cells in a physiological manner to mediate rapid and controlled T-cell expansion. APC-ms are composed of individual high-aspect-ratio silica microrods loaded with soluble mitogenic cues and coated with liposomes of defined compositions, to form supported lipid bilayers. Membrane-bound ligands for stimulation and co-stimulation of T-cell receptors are presented via the fluid, synthetic membranes, while mitogenic cues are released slowly from the microrods. In culture, interacting T cells assemble the individual APC-ms microrods into a biodegradable 3D matrix. Compared to conventional methods, APC-ms facilitates several-fold greater polyclonal T-cell expansion and improved antigen-specific enrichment of rare T-cell subpopulations. Here we provide a detailed protocol for APC-ms synthesis and use for human T-cell activation, and discuss important considerations for material design and T-cell co-culture. This protocol describes the facile assembly of APC-ms in ~4 h and rapid expansion or enrichment of relevant T-cell clones in <2 weeks, and is applicable for T-cell manufacturing and assay development. %B Nat Protoc %V 15 %P 773-798 %8 2020 03 %G eng %N 3 %1 http://www.ncbi.nlm.nih.gov/pubmed/31932772?dopt=Abstract %R 10.1038/s41596-019-0249-0