Abstract
T cells migrate diverse microenvironments of the body to mount antigen-specific immune responses. T cell activation, a key initial process for antigen-specific immune responses, occur in secondary lymphoid organs such as spleens and lymph nodes where high density of T cells migrates rapidly through the reticular networks formed by stromal cells. In vitro model system recapitulating key characteristics of secondary lymphoid organs, confined spaces densely packed with rapidly migrating cells, would be useful to investigate mechanisms of T cell migration. In this study, we devised a method to fabricate microchannels densely packed with T cells. Microchannel arrays with fixed height (4 μm) and length (1.5 mm) and various widths (15~80 μm) were fabricated in between trapezoid-shaped reservoirs that facilitated T cell sedimentation near microchannel entries. Microchannel surface chemistry and filling time were optimized to achieve high packing density (0.89) of T cell filling within microchannels. Particle image velocimetry (PIV) analysis method was employed to extract velocity field of microchannels densely packed with T cells. Using velocity field information, various motility parameters were further evaluated to quantitatively assess the effects of microchannel width and media tonicity on T cell motility within cell dense microenvironments.
Highlights
T cells migrate diverse microenvironments of the body to mount antigen-specific immune responses
T cell activation by T cell-antigen presenting cells (APCs) interactions in secondary lymphoid organs is a key event for the initiation of antigen-specific immune responses
Www.nature.com/scientificreports microenvironments in secondary lymphoid organs such as spleens and lymph nodes (LNs), where high density of lymphocytes forms segregated compartments and exerts rapid motility through the reticular network generated by stromal cells within the compartments[20,21]
Summary
T cells migrate diverse microenvironments of the body to mount antigen-specific immune responses. T cell activation, a key initial process for antigen-specific immune responses, occur in secondary lymphoid organs such as spleens and lymph nodes where high density of T cells migrates rapidly through the reticular networks formed by stromal cells. Dendritic cell migration in peripheral tissue[16], T cell motility in interstitial spaces regulated by myosin proteins[17,18], and leukocyte chemotactic responses[19] were studied using microchannel devices. This simple model has been extremely useful for mechanistic study because motility of leukocytes in microchannels was similar to that of in vivo interstitial spaces, whereas cell manipulation and data acquisition/processing are much easier than intravital imaging. We adjusted tonicity of media to study the role of cell membrane tension on T cell migration within microchannels densely packed with T cells
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