Mobile sensing based on the integration of microfluidic device and smartphone, so-called MS 2 technology, has enabled many applications over recent years, and continues to stimulate growing interest in both research communities and industries. In particular, it has been envisioned that MS 2 technology can be developed for various cell functional assays to enable basic research and clinical applications. Toward this direction, in this paper, we describe the development of a MS 2 -based cell functional assay for testing cell migration (the M kit ). The system is constructed as an integrated test kit, which includes microfluidic chips, a smartphone-based imaging platform, the phone apps for image capturing and data analysis, and a set of reagent and accessories for performing the cell migration assay. We demonstrated that the M kit can effectively measure purified neutrophil and cancer cell chemotaxis. Furthermore, neutrophil chemotaxis can be tested from a drop of whole blood using the M kit with red blood cell (RBC) lysis. The effects of chemoattractant dose and gradient profile on neutrophil chemotaxis were also tested using the M kit . In addition to research applications, we demonstrated the effective use of the M kit for on-site test at the hospital and for testing clinical samples from chronic obstructive pulmonary disease patient. Thus, this developed M kit provides an easy and integrated experimental platform for cell migration related research and potential medical diagnostic applications.
The importance of the bi-directional natural killer–dendritic cell crosstalk in coordinating anti-tumour and anti-microbial responses in vivo has been well established. However, physical parameters associated with natural killer–dendritic cell interactions have not been fully elucidated. We have previously used a simple “Y” shaped microfluidic device to study natural killer cell-migratory responses toward chemical gradients from a conditioned medium of dendritic cells. There are, however, limitations of the Y-shaped microfluidic devices that could not support higher throughput analyses and studies of cell–cell interactions. Here, we report two novel microfluidic devices (D3-Chip, T2-Chip) we applied in advanced studies of natural killer-cell migrations and their interactions with dendritic cells in vitro. The D3-Chip is an improved version of the previously published Y-shaped device that supports high-throughput analyses and docking of the cells of interest in the migration assay before they are exposed to a chemical gradient. The T2-Chip is created to support analyses of natural killer–dendritic cell cell–cell interactions without the requirement of promoting a natural killer cell to migrate long distances to find a loaded dendritic cell in the device. Using these two microfluidic platforms, we observe quantitative differences in the abilities of the immature and lipopolysaccharide-activated mature dendritic cells to interact with activated natural killer cells. The contact time between the activated natural killer cells and immature dendritic cells is significantly longer than that of the mature dendritic cells. There is a significantly higher frequency of an immature dendritic cell coming into contact with multiple natural killer cells and/or making multiple simultaneous contacts with multiple natural killer cells. To contrast, an activated natural killer cell has a significantly higher frequency of coming into contact with the mature dendritic cells than immature dendritic cells. Collectively, these differences in natural killer–dendritic cell interactions may underlie the differential maturation of immature dendritic cells by activated natural killer cells. Further applications of these microfluidic devices in studying natural killer–dendritic cell crosstalk under defined microenvironments shall enrich our understanding of the functional regulations of natural killer cells and dendritic cells in the natural killer–dendritic cell crosstalk.
The amoeboid-like cell motility is known to be driven by the acidic enzymatic hydrolysis of ATP in the actin-myosin system. However, the electro-mechano-chemical coupling, whereby the free energy of ATP hydrolysis is transformed into the power of electrically polarized cell movement, is poorly understood. Previous experimental studies showed that actin filaments motion, cytoplasmic streaming, and muscle contraction can be reconstituted under actin-activated ATP hydrolysis by soluble non-filamentous myosin fragments. Thus, biological motility was demonstrated in the absence of a continuous protein network. These results lead to an integrative conceptual model for cell motility, which advocates an active role played by intracellular proton currents and cytoplasmic streaming (iPC-CS). In this model, we propose that protons and fluid currents develop intracellular electric polarization and pressure gradients, which generate an electro-hydrodynamic mode of amoeboid motion. Such energetic proton currents and active streaming are considered to be mainly driven by stereospecific ATP hydrolysis through myosin heads along oriented actin filaments. Key predictions of this model are supported by microscopy visualization and in-depth sub-population analysis of purified human neutrophils using a microfluidic electrotaxis assay. Three distinct phases in cell motility profiles, morphology, and cytoplasmic streaming in response to physiological ranges of chemoattractant stimulation and electric field application are revealed. Our results support an intrinsic electric dipole formation linked to different patterns of cytoplasmic streaming, which can be explained by the iPC-CS model. Collectively, this alternative biophysical mechanism of cell motility provides new insights into bioenergetics with relevance to potential new biomedical applications.
Netrin‐1 is well‐known for its chemoattractive and chemorepulsive properties for axon guidance. Early studies report that netrin‐1 inhibits granulocyte migration. On the other hand, netrin‐1 can promote cancer cell migration and invasion. The underlying mechanisms are not well understood, which requires more in‐depth characterizations of netrin‐1 mediated immune and cancer cell migration. The present study, for the first time, employs microfluidic devices that are recently developed to quantitatively investigate the effects of netrin‐1 on the motility and chemotaxis of human blood neutrophils and human breast cancer cells under well‐controlled gradient conditions. The results show that netrin‐1 reduces chemokinetic motility of human neutrophils, which is accompanied with reduced cell polarization and spreading. In addition, netrin‐1 reduces neutrophil chemotaxis to N‐formyl‐Met‐Leu‐Phe on fibronectin substrate but interestingly not on collagen substrate. By contrast, netrin‐1 promotes the migration of human breast cancer cells. Furthermore, it is found that netrin‐1 reduces neutrophil chemotaxis to the supernatant of human breast cancer cell culture. Collectively, this microfluidic cell migration study provides quantitative characterizations of the effects of netrin‐1 on the motility and chemotaxis of neutrophils and breast cancer cells, and further suggests the potential role of netrin‐1 in regulating neutrophil recruitment to breast cancer microenvironments.
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