Continuous Cell Separation Using Microfluidic-Based Cell Retention Device with Alternative Boosted Flow.

The development of a continuous process for cell separation is growing rapidly due to the current trend of cost-effective manufacturing in biological industries. The continuous cell separation process has a significant reduction in capital equipment costs and facility size compared to the conventional batch process. In the study, a multi-layered microfluidic-based device integrated with the porous membranes was fabricated for continuous size-based isolation of the cells based on the mechanism of restrictive cross-flow filtration, allowing the biological sample entered in a single inlet of the device and separated into two outlet streams. One stream which contained the cells returned back to the original sample fluid, while another stream with conditioned medium only was collected for later applications. The membrane fouling issue was overcome by introducing the alternative flow rate consisted of a set of higher and lower flows. The device integrated with the controllable flow restriction allows to increase the permeate flow rate, and alternative boosted flow demonstrates the high permeate flow rate (0.3 mL/min), high cell viability (> 98%), and increase of cell concentration (48%). As a result, we believe that the microfluidic-based continuous cell separation system is a promising tool for downstream bioprocess.

Plug-and-Play In Vitro Metastasis System toward Recapitulating the Metastatic Cascade.

Microfluidic-based tumor models that mimic tumor culture environment have been developed to understand the cancer metastasis mechanism and discover effective antimetastatic drugs. These models successfully recapitulated key steps of metastatic cascades, yet still limited to few metastatic steps, operation difficulty, and small molecule absorption. In this study, we developed a metastasis system made of biocompatible and drug resistance plastics to recapitulate each metastasis stage in three-dimensional (3D) mono- and co-cultures formats, enabling the investigation of the metastatic responses of cancer cells (A549-GFP). The plug-and-play feature enhances the efficiency of the experimental setup and avoids initial culture failures. The results demonstrate that cancer cells tended to proliferate and migrate with circulating flow and intravasated across the porous membrane after a period of 3 d when they were treated with transforming growth factor-beta 1 (TGF-ß1) or co-cultured with human pulmonary microvascular endothelial cells (HPMECs). The cells were also observed to detach and migrate into the circulating flow after a period of 20 d, indicating that they transformed into circulating tumor cells for the next metastasis stage. We envision this metastasis system can provide novel insights that would aid in fully understanding the entire mechanism of tumor invasion.