Integrated Microfluidic System for Gene Silencing and Cell Migration

Abstract

Metastasis involves the phenotype transition of cancer cells to gain invasiveness, and the following migration at the tumor site. Here an integrated microfluidic chip to study this process is presented by combining on-chip delivery of siRNA for gene silencing and cell migration assay. The major advantage of the integrated chip is the simple input of cells and gene transfection materials, and the ultimate output of migration ability. The reverse-fishbone structure and 0.7× phosphate-buffered saline solution are the optimized parameters for improved delivery efficiency. Using the chip, it is validated that cofilin plays an essential role in regulating cancer cell migration. The integrated chip may provide a simple and effective platform for biologists to easily check the role of specific genes in metastasis.

Leveraging the elastic deformability of polydimethylsiloxane microfluidic channels for efficient intracellular delivery

“…5 μm constriction) sustained irreversible membrane damage that did not heal over time as compared to larger constriction sizes that did not induce an as extreme morphological deformation as the smallest constriction size. This observation is consistent with a previous report 19 where cells that underwent extreme membrane perturbations were less likely to remain viable. We also observed a small decrease in viability for the sample treated with the commercial transfection reagent, which was expected since lipid-based transfection reagents are known to have a slight cytotoxic effect on the treated cells, especially at long exposure times.…”

Leveraging the elastic deformability of polydimethylsiloxane microfluidic channels for efficient intracellular delivery

“…5 μm constriction) sustained irreversible membrane damage that did not heal over time as compared to larger constriction sizes that did not induce an as extreme morphological deformation as the smallest constriction size. This observation is consistent with a previous report 19 where cells that underwent extreme membrane perturbations were less likely to remain viable. We also observed a small decrease in viability for the sample treated with the commercial transfection reagent, which was expected since lipid-based transfection reagents are known to have a slight cytotoxic effect on the treated cells, especially at long exposure times.…”

Leveraging the Elastic Deformability of Polydimethylsiloxane Microfluidic Channels for Efficient Intracellular Delivery

“…5 µm constriction) sustained irreversible membrane damage that did not heal over time as compared to larger constriction sizes that did not induce an as extreme morphological deformation as the smallest constriction size. This observation is consistent with [19] where cells that underwent extreme membrane perturbations were less likely to remain viable. We also observed a small decrease in viability for the sample treated with the commercial transfection reagent, which was expected since lipid-based transfection reagents are known to have a slight cytotoxic effect on the treated cells, especially at long exposure times [30] .…”

“…Liquid flow forces the cell to traverse further through the constriction where its morphology begins to flatten to match the clearance between the tip of the micro-feature and the opposite inner wall of the channel. This rapid deformation has been previously shown to result in the creation of transient pores through the cellular membrane [15, 19-21, 29] . Passive diffusion of the desired cargo occurs rapidly at this stage from the extracellular media surrounding the cell along the concentration gradient.…”

Intracellular Delivery by Membrane Disruption: Mechanisms, Strategies, and Concepts

“…(ii) COMSOL modeling indicates the stress (N m –2 ) that the cell membrane would undergo upon passage through the different shapes of constrictions. Experiments and modeling show the reverse wishbone pattern as the most effective for localized membrane disruption in this device . Figure reprinted from ref .…”

“…Experiments and modeling show the reverse wishbone pattern as the most effective for localized membrane disruption in this device . Figure reprinted from ref . Copyright 2017, with permission from John Wiley and Sons.…”

Leveraging the elastic deformability of polydimethylsiloxane microfluidic channels for efficient intracellular delivery

“…5 μm constriction) sustained irreversible membrane damage that did not heal over time as compared to larger constriction sizes that did not induce an as extreme morphological deformation as the smallest constriction size. This observation is consistent with a previous report 19 where cells that underwent extreme membrane perturbations were less likely to remain viable. We also observed a small decrease in viability for the sample treated with the commercial transfection reagent, which was expected since lipid-based transfection reagents are known to have a slight cytotoxic effect on the treated cells, especially at long exposure times.…”

Leveraging the Elastic Deformability of Polydimethylsiloxane Microfluidic Channels for Efficient Intracellular Delivery

“…5 µm constriction) sustained irreversible membrane damage that did not heal over time as compared to larger constriction sizes that did not induce an as extreme morphological deformation as the smallest constriction size. This observation is consistent with [19] where cells that underwent extreme membrane perturbations were less likely to remain viable. We also observed a small decrease in viability for the sample treated with the commercial transfection reagent, which was expected since lipid-based transfection reagents are known to have a slight cytotoxic effect on the treated cells, especially at long exposure times [30] .…”

“…Liquid flow forces the cell to traverse further through the constriction where its morphology begins to flatten to match the clearance between the tip of the micro-feature and the opposite inner wall of the channel. This rapid deformation has been previously shown to result in the creation of transient pores through the cellular membrane [15, 19-21, 29] . Passive diffusion of the desired cargo occurs rapidly at this stage from the extracellular media surrounding the cell along the concentration gradient.…”

Intracellular Delivery by Membrane Disruption: Mechanisms, Strategies, and Concepts

“…(ii) COMSOL modeling indicates the stress (N m –2 ) that the cell membrane would undergo upon passage through the different shapes of constrictions. Experiments and modeling show the reverse wishbone pattern as the most effective for localized membrane disruption in this device . Figure reprinted from ref .…”

“…Experiments and modeling show the reverse wishbone pattern as the most effective for localized membrane disruption in this device . Figure reprinted from ref . Copyright 2017, with permission from John Wiley and Sons.…”

Leveraging the elastic deformability of polydimethylsiloxane microfluidic channels for efficient intracellular delivery

“…5 μm constriction) sustained irreversible membrane damage that did not heal over time as compared to larger constriction sizes that did not induce an as extreme morphological deformation as the smallest constriction size. This observation is consistent with a previous report 19 where cells that underwent extreme membrane perturbations were less likely to remain viable. We also observed a small decrease in viability for the sample treated with the commercial transfection reagent, which was expected since lipid-based transfection reagents are known to have a slight cytotoxic effect on the treated cells, especially at long exposure times.…”

Leveraging the Elastic Deformability of Polydimethylsiloxane Microfluidic Channels for Efficient Intracellular Delivery

“…5 µm constriction) sustained irreversible membrane damage that did not heal over time as compared to larger constriction sizes that did not induce an as extreme morphological deformation as the smallest constriction size. This observation is consistent with [19] where cells that underwent extreme membrane perturbations were less likely to remain viable. We also observed a small decrease in viability for the sample treated with the commercial transfection reagent, which was expected since lipid-based transfection reagents are known to have a slight cytotoxic effect on the treated cells, especially at long exposure times [30] .…”

“…Liquid flow forces the cell to traverse further through the constriction where its morphology begins to flatten to match the clearance between the tip of the micro-feature and the opposite inner wall of the channel. This rapid deformation has been previously shown to result in the creation of transient pores through the cellular membrane [15, 19-21, 29] . Passive diffusion of the desired cargo occurs rapidly at this stage from the extracellular media surrounding the cell along the concentration gradient.…”

Intracellular Delivery by Membrane Disruption: Mechanisms, Strategies, and Concepts

“…(ii) COMSOL modeling indicates the stress (N m –2 ) that the cell membrane would undergo upon passage through the different shapes of constrictions. Experiments and modeling show the reverse wishbone pattern as the most effective for localized membrane disruption in this device . Figure reprinted from ref .…”

“…Experiments and modeling show the reverse wishbone pattern as the most effective for localized membrane disruption in this device . Figure reprinted from ref . Copyright 2017, with permission from John Wiley and Sons.…”