Longitudinal multiparameter assay of lymphocyte interactions from onset by microfluidic cell pairing and culture
Resolving how the early signaling events initiated by cell-cell interactions are transduced into diverse functional outcomes necessitates correlated measurements at various stages. Typical approaches that rely on bulk cocultures and population-wide correlations, however, only reveal these relationsh...
| Main Authors: | , , , , , |
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| Other Authors: | , , , , , |
| Format: | Article |
| Language: | English |
| Published: |
National Academy of Sciences (U.S.),
2016-12-27T21:43:00Z.
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| Subjects: | |
| Online Access: | Get fulltext |
| Summary: | Resolving how the early signaling events initiated by cell-cell interactions are transduced into diverse functional outcomes necessitates correlated measurements at various stages. Typical approaches that rely on bulk cocultures and population-wide correlations, however, only reveal these relationships broadly at the population level, not within each individual cell. Here, we present a microfluidics-based cell-cell interaction assay that enables longitudinal investigation of lymphocyte interactions at the single-cell level through microfluidic cell pairing, on-chip culture, and multiparameter assays, and allows recovery of desired cell pairs by micromanipulation for off-chip culture and analyses. Well-defined initiation of interactions enables probing cellular responses from the very onset, permitting single-cell correlation analyses between early signaling dynamics and later-stage functional outcomes within same cells. We demonstrate the utility of this microfluidic assay with natural killer cells interacting with tumor cells, and our findings suggest a possible role for the strength of early calcium signaling in selective coordination of subsequent cytotoxicity and IFN-gamma production. Collectively, our experiments demonstrate that this new approach is well-suited for resolving the relationships between complex immune responses within each individual cell. Singapore-MIT Alliance American Association for Cancer Research. Pancreatic Cancer Action Network Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science (Frank Quick Faculty Research Innovation Fellowship) |
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