A droplet‐merging platform for comparative functional analysis of m1 and m2 macrophages in response to <i>e. coli</i>‐induced stimuli

Hondroulis, Evangelia; Movila, Alexandru; Sabhachandani, Pooja; Sarkar, Saheli; Cohen, Noa; Kawai, Toshihisa; Konry, Tania

doi:10.1002/bit.26196

Cited by 13 publications

(11 citation statements)

References 20 publications

(40 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We developed a droplet-based cytotoxicity assay to quantify features of dynamic cell–cell interactions and determine NK cell-mediated cytolysis of target cells at single-cell level. CD56 + NK cells were co-encapsulated with target lymphoma cells in picoliter-volume droplets in an integrated microfluidic docking array ( 31 , 46 ). NK cells were incubated through one inlet of the device and target lymphoma cells through the other inlet in a single-step cell loading process (Figure 2 A).…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Dynamic Analysis of Human Natural Killer Cell Response at Single-Cell Resolution in B-Cell Non-Hodgkin Lymphoma

et al. 2017

Self Cite

View full text Add to dashboard Cite

Natural killer (NK) cells are phenotypically and functionally diverse lymphocytes that recognize and kill cancer cells. The susceptibility of target cancer cells to NK cell-mediated cytotoxicity depends on the strength and balance of regulatory (activating/inhibitory) ligands expressed on target cell surface. We performed gene expression arrays to determine patterns of NK cell ligands associated with B-cell non-Hodgkin lymphoma (b-NHL). Microarray analyses revealed significant upregulation of a multitude of NK-activating and costimulatory ligands across varied b-NHL cell lines and primary lymphoma cells, including ULBP1, CD72, CD48, and SLAMF6. To correlate genetic signatures with functional anti-lymphoma activity, we developed a dynamic and quantitative cytotoxicity assay in an integrated microfluidic droplet generation and docking array. Individual NK cells and target lymphoma cells were co-encapsulated in picoliter-volume droplets to facilitate monitoring of transient cellular interactions and NK cell effector outcomes at single-cell level. We identified significant variability in NK-lymphoma cell contact duration, frequency, and subsequent cytolysis. Death of lymphoma cells undergoing single contact with NK cells occurred faster than cells that made multiple short contacts. NK cells also killed target cells in droplets via contact-independent mechanisms that partially relied on calcium-dependent processes and perforin secretion, but not on cytokines (interferon-γ or tumor necrosis factor-α). We extended this technique to characterize functional heterogeneity in cytolysis of primary cells from b-NHL patients. Tumor cells from two diffuse large B-cell lymphoma patients showed similar contact durations with NK cells; primary Burkitt lymphoma cells made longer contacts and were lysed at later times. We also tested the cytotoxic efficacy of NK-92, a continuously growing NK cell line being investigated as an antitumor therapy, using our droplet-based bioassay. NK-92 cells were found to be more efficient in killing b-NHL cells compared with primary NK cells, requiring shorter contacts for faster killing activity. Taken together, our combined genetic and microfluidic analysis demonstrate b-NHL cell sensitivity to NK cell-based cytotoxicity, which was associated with significant heterogeneity in the dynamic interaction at single-cell level.

show abstract

Section: Resultsmentioning

confidence: 99%

“…The microfluidic devices were fabricated by standard soft-lithography protocols ( 31 , 44 – 46 ). The device design was formulated using CAD (CAD/Art Services, Bandon, OR, USA) and printed on a transparency photomask (Fine Line Imaging, Colorado Springs, CO, USA).…”

Section: Methodsmentioning

confidence: 99%

Dynamic Analysis of Human Natural Killer Cell Response at Single-Cell Resolution in B-Cell Non-Hodgkin Lymphoma

et al. 2017

Self Cite

View full text Add to dashboard Cite

show abstract

“…Dynamics of CD8+ T cell mediated cytolysis of myeloma cell lines was also monitored in the same work (Figure d) . In another work of the same group, they co‐encapsulated polarized M1 and M2 macrophages with Escherichia coli ( E. coli ) in the same platform . Phagocytic activity of M1 and M2 macrophages was determined by loss of E. coli fluorescence and cell counts in the co‐encapsulated droplets.…”

Section: Monitoring Of Immune Cell Cytotoxicity On a Chipmentioning

confidence: 89%

Biochips—New Platforms for Cell‐Based Immunological Assays

Shao

Qin

2017

Small Methods

View full text Add to dashboard Cite

Considerable advances have been witnessed in the development of biochips that seek to realize various types of immune cell analysis on microscale platforms and enhance both basic and applied immunological research beyond the capability of conventional methods. Here, state‐of‐the‐art designs and examples for biochip‐based analysis and manipulation of immune cells are reviewed, and the potential of this emerging technology to enhance the understanding of immunology and improve disease diagnosis and treatment is discussed. In particular, some of the recent advances in this field, along with the challenges that must be addressed for these technologies, and their potential in precision medicine are highlighted.

show abstract

“…This structure could also be used to merge cells and interested constituents, such as drugs and nutrients, into single droplets, proceeding phenotypic constituent profiling. 82 Hondroulis et al 83 reported an approach based on the docking platform to co-encapsulate polarized macrophages with Escherichia coli, facilitating functional analysis of macrophage heterogeneity quantitatively and living cell profiling of effector immune cells in situ. Dynamic analysis of human natural killer cell response with B lymphocyte in B-cell non-Hodgkin lymphoma was reported using this merging platform as well.…”

Section: Cell Culturementioning

confidence: 99%