Utilizing Custom-designed Galvanotaxis Chambers to Study Directional Migration of Prostate Cells

Yang, Hsin-Ya; La, Thi Dinh; Isseroff, Roslyn Rivkah

doi:10.3791/51973

Cited by 7 publications

(3 citation statements)

References 34 publications

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“…Cells were resuspended in modified Hank’s Balanced solution (mHBSS, Sigma-Aldrich) with 1 μM N-Formyl-Met-Leu-Phe (fMLP, Sigma Aldrich), and then were then plated in a 3D-printed electrotaxis chamber composed of polylactic acid ( Yang et al, 2014 ). Toxicity and changes in pH due to electrode byproducts were minimized by using separate baths for each AgCl electrode and the cell-mHBSS solution, and by connecting the baths using agar bridges in mHBSS (2%, 0.5 cm diameter, created with glass tubing).…”

Section: Methodsmentioning

confidence: 99%

Actin Dynamics as a Multiscale Integrator of Cellular Guidance Cues

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Campanello

Hourwitz

et al. 2022

Front. Cell Dev. Biol.

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Migrating cells must integrate multiple, competing external guidance cues. However, it is not well understood how cells prioritize among these cues. We investigate external cue integration by monitoring the response of wave-like, actin-polymerization dynamics, the driver of cell motility, to combinations of nanotopographies and electric fields in neutrophil-like cells. The electric fields provide a global guidance cue, and approximate conditions at wound sites in vivo. The nanotopographies have dimensions similar to those of collagen fibers, and act as a local esotactic guidance cue. We find that cells prioritize guidance cues, with electric fields dominating long-term motility by introducing a unidirectional bias in the locations at which actin waves nucleate. That bias competes successfully with the wave guidance provided by the bidirectional nanotopographies.

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Section: Methodsmentioning

confidence: 99%

Actin Dynamics as a Multiscale Integrator of Cellular Guidance Cues

Bull

Campanello

Hourwitz

et al. 2022

Front. Cell Dev. Biol.

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show abstract

“…Cells were resuspended in modified Hank's Balanced solution (mHBSS, Sigma-Aldrich) with 1 µM N-Formyl-Met-Leu-Phe (fMLP, Sigma Aldrich), and then were then plated in a 3D-printed electrotaxis chamber composed of polylactic acid (38). Toxicity and changes in pH due to electrode byproducts were minimized by using separate baths for each AgCl electrode and the cell-mHBSS solution, and by connecting the baths using agar bridges in mHBSS (2%, 0.5 cm diameter, created with glass tubing).…”

Section: Methodsmentioning

confidence: 99%

Actin dynamics as a multiscale integrator of cellular guidance cues

Bull

Campanello

et al. 2021

Preprint

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Cells are able to integrate multiple, and potentially competing, cues to determine a migration direction. For instance, in wound healing, cells follow chemical signals or electric fields to reach the wound edge, regardless of any local guidance cues. To investigate this integration of guidance cues, we monitor the actin-polymerization dynamics of immune cells in response to cues on a subcellular scale (nanotopography) and on the cellular scale (electric fields, EFs). In the fast, amoeboid-type migration, commonly observed in immune cells, actin polymerization at the cell's leading edge is the driver of motion. The excitable systems character of actin polymerization leads to self-propagating, two-dimensional wavefronts that enable persistent cell motion. We show that EFs guide these wavefronts, leading to turning of cells when the direction of the EF changes. When nanoridges promote one-dimensional (1D) waves of actin polymerization that move along the ridges (esotaxis), EF guidance along that direction is amplified. 1D actin waves cannot turn or change direction, so cells respond to a change in EF direction by generating new 1D actin waves. At the cellular scale, the emergent response is a turning of the cell. For nanoridges perpendicular to the direction of the EF, the 1D actin waves are guided by the nanotopography, but both the average location of new actin waves and the whole cell motion are guided by the EF. Thus, actin waves respond to each cue on its intrinsic length scale, allowing cells to exhibit versatile responses to the physical microenvironment.

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“…Electrotaxis is a guiding mechanism for the orientation and directional movements of many cell types, including fission yeast cells (12), Caenorhabditis elegans (13)(14)(15)(16), pathogenic bacterium such as Pseudomonas aeruginosa, Escherichia coli, and Dictyostelium (17)(18)(19)(20). It has been reported that some cells migrate toward the cathode [e.g., neural stem cells (21)(22)(23)(24)(25)(26), fibroblasts (27)(28)(29), keratinocytes (30)(31)(32)(33), rat prostate cancer cells (34,35), T lymphocyte (36)(37)(38)(39)(40), lung cancer cells (41)(42)(43)(44)(45), and many epithelial cell types (3,30,(46)(47)(48)(49)(50)(51)], and other cells migrate to the anode [e.g., corneal endothelial cells (52), breast cancer cells (53,54), glioblastoma (55,…”

Section: Introductionmentioning

confidence: 99%

Electrotaxis-on-Chip to Quantify Neutrophil Migration Towards Electrochemical Gradients

et al. 2021

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Electric fields are generated in vivo in a variety of physiologic and pathologic settings, including wound healing and immune response to injuries to epithelial barriers (e.g. lung pneumocytes). Immune cells are known to migrate towards both chemical (chemotaxis), physical (mechanotaxis) and electric stimuli (electrotaxis). Electrotaxis is the guided migration of cells along electric fields, and has previously been reported in T-cells and cancer cells. However, there remains a need for engineering tools with high spatial and temporal resolution to quantify EF guided migration. Here we report the development of an electrotaxis-on-chip (ETOC) platform that enables the quantification of dHL-60 cell, a model neutrophil-like cell line, migration toward both electrical and chemoattractant gradients. Neutrophils are the most abundant white blood cells and set the stage for the magnitude of the immune response. Therefore, developing engineering tools to direct neutrophil migration patterns has applications in both infectious disease and inflammatory disorders. The ETOC developed in this study has embedded electrodes and four migration zones connected to a central cell-loading chamber with migration channels [10 µm X 10 µm]. This device enables both parallel and competing chemoattractant and electric fields. We use our novel ETOC platform to investigate dHL-60 cell migration in three biologically relevant conditions: 1) in a DC electric field; 2) parallel chemical gradient and electric fields; and 3) perpendicular chemical gradient and electric field. In this study we used differentiated leukemia cancer cells (dHL60 cells), an accepted model for human peripheral blood neutrophils. We first quantified effects of electric field intensities (0.4V/cm-1V/cm) on dHL-60 cell electrotaxis. Our results show optimal migration at 0.6 V/cm. In the second scenario, we tested whether it was possible to increase dHL-60 cell migration to a bacterial signal [N-formylated peptides (fMLP)] by adding a parallel electric field. Our results show that there was significant increase (6-fold increase) in dHL60 migration toward fMLP and cathode of DC electric field (0.6V/cm, n=4, p-value<0.005) vs. fMLP alone. Finally, we evaluated whether we could decrease or re-direct dHL-60 cell migration away from an inflammatory signal [leukotriene B4 (LTB4)]. The perpendicular electric field significantly decreased migration (2.9-fold decrease) of dHL60s toward LTB4vs. LTB4 alone. Our microfluidic device enabled us to quantify single-cell electrotaxis velocity (7.9 µm/min ± 3.6). The magnitude and direction of the electric field can be more precisely and quickly changed than most other guidance cues such as chemical cues in clinical investigation. A better understanding of EF guided cell migration will enable the development of new EF-based treatments to precisely direct immune cell migration for wound care, infection, and other inflammatory disorders.

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Utilizing Custom-designed Galvanotaxis Chambers to Study Directional Migration of Prostate Cells

Cited by 7 publications

References 34 publications

Actin Dynamics as a Multiscale Integrator of Cellular Guidance Cues

Actin Dynamics as a Multiscale Integrator of Cellular Guidance Cues

Actin dynamics as a multiscale integrator of cellular guidance cues

Electrotaxis-on-Chip to Quantify Neutrophil Migration Towards Electrochemical Gradients

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