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Boukany, P. E.

Publications and source records attributed to Boukany, P. E..

5 recordsLinked to original sources

Label-free Isolation of Heterogeneous Breast Cancer Cell Populations via Insulator-Based Dielectrophoresis

Circulating tumor cells (CTCs) are valuable biomarkers for cancer diagnosis and monitoring, yet their isolation from blood remains challenging due to their phenotypic heterogeneity and rarity. Label-free microfluidic technologies offer a promising alternative to affinity-based approaches by exploiting intrinsic biophysical differences between cell types. Here, we developed a microfluidic platform for label-free cell separation based on insulator-based dielectrophoresis (iDEP). The microfluidic device employs an array of triangular insulating structures that generate strong electric field gradients in response to an externally applied alternating current (AC) electric field, enabling selective isolation of breast cancer cells from blood cells based on their dielectric properties. Hydrodynamic focusing is used to confine the sample stream and precisely control cell trajectories within the separation region. Numerical simulations were performed to optimize the electric field distribution and fluid flow characteristics within the device. Experimental validation using breast cancer cell lines (mesenchymal-like MDA-MB-231 cells and epithelial-like MCF-7 cells) spiked into peripheral blood mononuclear cells (PBMCs) demonstrates selective dielectrophoretic deflection of cancer cells while PBMCs largely follow the central streamline. The platform achieves recovery rates exceeding 98% and a separation purity above 65% within the optimized operating conditions. The proposed system provides a simple label-free approach to separate heterogeneous cell populations and represents a promising tool for microfluidic liquid biopsy enrichment applications.

cancer biology↗

Septins promote breast cancer cell invasion in 3D collagen gels by influencing actin-based protrusion formation

Septins are cytoskeletal proteins that contribute to essential cellular processes such as cell migration and cell division through interactions with the cell membrane and the cytoskeleton. High expression of septins is correlated with breast cancer malignancy and promotes cell invasion, but the molecular complexity of septins interactions has made it challenging to dissect the underlying molecular mechanisms. Here, we used a conditional knockout approach to deplete SEPT7 in the metastatic triple-negative breast cancer cell line Hs578T and examined the role of septin in 3D-matrix invasion of breast cancer cells. We show by spheroid assays that SEPT7 deletion strongly impairs breast cancer cell invasion into collagen gels. Additional single-cell migration studies using 3D collagen gels and microfluidic pillar devices that mimic the pores present in collagen matrices showed that SEPT7 expression regulates confined cell migration through control of cell shape and actin-based protrusions.

cancer biology↗

Cancer cell deformability impacts the rate of confined migration but not decision making

Cancer cells can utilize different invasion strategies to overcome physical arrest during confined migration through tissues with small pores. Cancer cell plasticity allows switches between different migration modes and transitions between single-cell and collective migration. The biophysical parameters that guide these decisions are poorly understood. In this work we investigated the link between cell deformability and migration efficacy in constrictions of two mesenchymal cancer cell types with similar invasion strategies: HT1080 fibrosarcoma cells and MV3 melanoma cells. To this end, we designed microfluidic platforms for (1) high-throughput cell deformability measurements and (2) migration through a variety of confining geometries. We measured different deformabilities for HT1080 and MV3 cells and correlated this to their migration efficacy through confinements. However, higher deformability and improved squeezing ability did not impact decision-making at junctions of channels of different widths. Our findings show that cell deformability correlates with better squeezing abilities through confinements, but does not impact directionality decisions.

cancer biology↗

Invasive cancer cells soften collagen networks and disrupt stress-stiffening via volume exclusion, contractility and adhesion

Collagen networks form the structural backbone of the extracellular matrix in both healthy and cancerous tissues, exhibiting nonlinear mechanical properties that crucially regulate tissue mechanics and cell behavior. Here, we investigate how the presence of invasive breast cancer cells (MDA-MB-231) influences the polymerization kinetics and mechanics of collagen networks using bulk shear rheology and rheo-confocal microscopy. We show that embedded cancer cells delay the onset of collagen polymerization due to volume exclusion effects. During polymerization, the cells (at 4% volume fraction) cause an unexpected time-dependent softening of the network. We show that this softening effect arises from active remodeling via adhesion and contractility rather than from proteolytic degradation. At higher cell volume fractions, the dominant effect of the cells shifts to volume exclusion, causing a two-fold reduction of network stiffness. Additionally, we demonstrate that cancer cells suppress the characteristic stress-stiffening response of collagen. This effect (partially) disappears when cell adhesion and contractility are inhibited, and it is absent when the cells are replaced by passive hydrogel particles. These findings provide new insights into how active inclusions modify the mechanics of fibrous networks, contributing to a better understanding of the role of cells in the mechanics of healthy and diseased tissues like invasive tumors.

biophysics↗

Intracellular transport of electrotransferred DNA cargo is governed by coexisting ergodic and non ergodic anomalous diffusion

The ability of exogenous DNA cargo to overcome the active and viscoelastic eukaryotic cytoplasm is a principal determinant for the gene delivery efficacy. During DNA electrotransfer, DNA forms complexes with the membrane (DNA cargo) which is transported through the cytoplasm through a combination of passive diffusion and active transport. However, this process is poorly understood limiting rational optimization of DNA cargo to be delivered to different cell types. We have investigated the intracellular transport of DNA cargo (of sizes 100 bp, 250 bp and 500 bp) delivered by electrotransfer to non-cancerous and cancerous mammalian cells. We demonstrate that intracellular DNA cargo transport is governed by coexisting ergodic and non ergodic anomalous diffusion for all the tested DNA sizes and cell types. The apparent diffusion coefficient of the electrotransferred DNA cargo in the cytoplasm decreases when the DNA size is increased from 100 bp to 500 bp. Interestingly, the electrotransferred DNA cargo (500 bp) transport is strongly dependent on the cells cancer state. Intracellular electrotransferred DNA cargo transport has a higher probability of superdiffusive transport and lower probability of caging in metastatic cells compared to malignant cells followed by benign cells.

biophysics↗