Longitudinal multiparameter assay of lymphocyte interactions from onset by microfluidic cell pairing and culture

作者：Dura Burak^*; Servos Mariah M; Barry Rachel M; Ploegh Hidde L; Dougan Stephanie K; Voldman Joel^*

来源：Proceedings of the National Academy of Sciences of the United States of America, 2016, 113(26): E3599-E3608.

DOI：10.1073/pnas.1515364113

• microfluidics
• cell pairing
• single-cell analysis
• cell-cell interactions
• multiparameter assay

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.

• 出版日期2016-6-28
• 单位MIT