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The Continuous Concentration of Particles and Cancer Cell Line Using Cell Margination in a Groove-Based Channel.

ABSTRACT: In the capillary venules, blood cells auto-separate with red blood cells aggregating near the centre of vessel and the nucleated cells marginating toward the wall of vessel. In this experiment, we used cell margination to help enrich the Jurkat cells via a groove-based channel which provides a vertical expansion-contraction structure, wherein the red blood cells invade the grooves and push the Jurkat cells to the bottom of the channel. The secondary flows induced by the anisotropic grooves bring the Jurkat cells to the right sidewall. Rigid, 13-µm diameter polystyrene particles were spiked into the whole blood to verify the operating principle under various working conditions, and then tests were carried out using Jurkat cells (~15 µm). The performance of this device was quantified by analysing the cell distribution in a transverse direction at the outlet, and then measuring the cell concentration from the corresponding outlets. The results indicate that Jurkat cells were enriched by 22.3-fold with a recovery rate of 83.4%, thus proving that this microfluidic platform provides a gentle and passive way to isolate intact and viable Jurkat cells.

SUBMITTER: Yan S

PROVIDER: S-EPMC6189968 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Chikungunya virus-like particles (VLPs) have potential to be used as a prophylactic vaccine based on testing in multiple animal models and are currently being evaluated for human use in a Phase I clinical trial. The current method for producing these enveloped alphavirus VLPs by transient gene expression in mammalian cells presents challenges for scalable and robust industrial manufacturing, so the insect cell baculovirus expression vector system was evaluated as an alternative expression technology. Subsequent to recombinant baculovirus infection of Sf21 cells in standard culture media (pH 6.2-6.4), properly processed Chikungunya structural proteins were detected and assembled capsids were observed. However, an increase in culture pH to 6.6-6.8 was necessary to produce detectable concentrations of assembled VLPs. Since this elevated production pH exceeds the optimum for growth medium stability and Sf21 culture, medium modifications were made and a novel insect cell variant (SfBasic) was derived by exposure of Sf21 to elevated culture pH for a prolonged period of time. The high-pH adapted SfBasic insect cell line described herein is capable of maintaining normal cell growth into the typical mammalian cell culture pH range of 7.0-7.2 and produces 11-fold higher Chikungunya VLP yields relative to the parental Sf21 cell line. After scale-up into stirred tank bioreactors, SfBasic derived VLPs were chromatographically purified and shown to be similar in size and structure to a VLP standard derived from transient gene expression in HEK293 cells. Total serum anti-Chikungunya IgG and neutralizing titers from guinea pigs vaccinated with SfBasic derived VLPs or HEK293 derived VLPs were not significantly different with respect to production method, suggesting that this adapted insect cell line and production process could be useful for manufacturing Chikungunya VLPs for use as a vaccine. The adaptation of Sf21 to produce high levels of recombinant protein and VLPs in an elevated pH range may also have applications for other pH-sensitive protein or VLP targets.

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The Continuous Concentration of Particles and Cancer Cell Line Using Cell Margination in a Groove-Based Channel.

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