Issue 15, 2016

Highly efficient adenoviral transduction of pancreatic islets using a microfluidic device

Abstract

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 ∼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.

Graphical abstract: Highly efficient adenoviral transduction of pancreatic islets using a microfluidic device

Supplementary files

Article information

Article type
Paper
Submitted
12 Mar 2016
Accepted
25 Jun 2016
First published
27 Jun 2016

Lab Chip, 2016,16, 2921-2934

Author version available

Highly efficient adenoviral transduction of pancreatic islets using a microfluidic device

P. N. Silva, Z. Atto, R. Regeenes, U. Tufa, Y. Y. Chen, W. C. W. Chan, A. Volchuk, D. M. Kilkenny and J. V. Rocheleau, Lab Chip, 2016, 16, 2921 DOI: 10.1039/C6LC00345A

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