In this paper, we report a new method to incorporate 3D scaffold with electrotaxis measurement in the microfluidic device. The electrotactic response of lung cancer cells in the 3D foam scaffolds which resemble the in vivo pulmonary alveoli may give more insight on cellular behaviors in vivo. The 3D scaffold consists of ordered arrays of uniform spherical pores in gelatin. We found that cell morphology in the 3D scaffold was different from that in 2D substrate. Next, we applied a direct current electric field (EF) of 338 mV/mm through the scaffold for the study of cells' migration within. We measured the migration directedness and speed of different lung cancer cell lines, CL1-0, CL1-5, and A549, and compared with those examined in 2D gelatin-coated and bare substrates. The migration direction is the same for all conditions but there are clear differences in cell morphology, directedness, and migration speed under EF. Our results demonstrate cell migration under EF is different in 2D and 3D environments and possibly due to different cell morphology and/or substrate

We report a new design of microfluidic chip (Multiple electric Field with Uniform Flow chip, MFUF chip) to create multiple electric field strengths (EFSs) while providing a uniform flow field simultaneously. MFUF chip was fabricated from poly-methyl methacrylates (PMMA) substrates by using CO2 laser micromachining. A microfluidic network with interconnecting segments was utilized to de-couple the flow field and the electric field (EF). Using our special design, different EFSs were obtained in channel segments that had an identical cross-section and therefore a uniform flow field. Four electric fields with EFS ratio of 7.9:2.8:1:0 were obtained with flow velocity variation of only 7.8% CV (coefficient of variation). Possible biological effect of shear force can therefore be avoided. Cell behavior under three EFSs and the control condition, where there is no EF, was observed in a single experiment. We validated MFUF chip performance using lung adenocarcinoma cell lines and then used the chip to study the electrotaxis of HSC-3, an oral squamous cell carcinoma cell line. The MFUF chip has high throughput capability for studying the EF-induced cell behavior under various EFSs, including the control condition (EFS = 0).

The wound-healing assay is an easy and economical way to quantify cell migration under diverse stimuli. Traditional assays such as scratch assays and barrier assays are widely and commonly used, but neither of them can represent the complicated condition when a wound occurs. It has been suggested that wound-healing is related to electric fields, which were found to regulate wound re-epithelialization. As a wound occurs, the disruption of epithelial barrier shortcircuits the trans-epithelial potential and then a lateral endogenous electric field is created. This field has been proved in vitro as an important cue for guiding the migration of fibroblasts, macrophages, and keratinocytes, a phenomenon termed electrotaxis or galvanotaxis. In this paper, we report a microfluidic electricalstimulated wound-healing chip (ESWHC) integrating electric field with a modified barrier assay. This chip was used to study the migration of fibroblasts under different conditions such as serum, electric field, and wound-healingpromoting drugs. We successfully demonstrate the feasibility of ESWHC to effectively and quantitatively study cell migration during wound-healing process, and therefore this chip could be useful in drug discovery and drug safety tests.