Journal
LAB ON A CHIP
Volume 16, Issue 15, Pages 2921-2934Publisher
ROYAL SOC CHEMISTRY
DOI: 10.1039/c6lc00345a
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Funding
- NSERC Discovery [RGPIN371705]
- NSERC Engage Grant
- CFI Leaders Opportunity Fund Grant
- BBDC Sun Life Pilot Grant
- NSERC [2015-06397]
- CIHR [MOP-130143, RMF-111623]
- NSERC PGS D awards
- UHN Graduate Award through University of Toronto Faculty of Medicine Banting
- Best Diabetes Centre (PNS)
- Charles Hollenberg Summer Studentship through Banting and Best Diabetes Centre
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Tissues are challenging to genetically manipulate due to limited penetration of viral particles resulting in low transduction efficiency. We are particularly interested in expressing genetically-encoded sensors in ex vivo pancreatic islets to measure glucose-stimulated metabolism, however poor viral penetration biases these measurements to only a subset of cells at the periphery. To increase mass transfer of viral particles, we designed a microfluidic device that holds islets in parallel hydrodynamic traps connected by an expanding by-pass channel. We modeled viral particle flow into the tissue using fluorescently-labelled gold nanoparticles of varying sizes and showed a penetration threshold of only similar to 5 nm. To increase this threshold, we used EDTA to transiently reduce cell-cell adhesion and expand intercellular space. Ultimately, a combination of media flow and ETDA treatment significantly increased adenoviral transduction to the core of the islet. As proof-of-principle, we used this protocol to transduce an ER-targeted redox sensitive sensor (eroGFP), and revealed significantly greater ER redox capacity at core islet cells. Overall, these data demonstrate a robust method to enhance transduction efficiency of islets, and potentially other tissues, by using a combination of microfluidic flow and transient tissue expansion.
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