细胞生物学
嵌合抗原受体
抗原提呈细胞
细胞
细胞培养
细胞疗法
离体
抗原
T细胞
分子生物学
化学
生物
免疫学
体外
干细胞
生物化学
免疫系统
遗传学
作者
David Zhang,Alexander S. Cheung,David J. Mooney
出处
期刊:Nature Protocols
[Springer Nature]
日期:2020-01-13
卷期号:15 (3): 773-798
被引量:45
标识
DOI:10.1038/s41596-019-0249-0
摘要
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.
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