TY - JOUR
T1 - On Demand Release and Retrieval of Bacteria from Microwell Arrays Using Photodegradable Hydrogel Membranes
AU - Van Der Vlies, André J.
AU - Barua, Niloy
AU - Nieves-Otero, Priscila A.
AU - Platt, Thomas G.
AU - Hansen, Ryan R.
N1 - Funding Information:
We acknowledge support from the National Science Foundation under Grant 1650187 and Kansas State University. P.A.N. acknowledges support from the National Science Foundation Graduate Research Fellowship under Grant No. GGVP004842. We would like to thank Dr. Urara Hasegawa (Chemical Engineering Department, Kansas State University) for help with confocal microscopy and use of her equipment for chemical synthesis, Dr. Alvaro Herrera (Biomolecular NMR Facility, Kansas State University) for measuring H NMR, Prof. Douglas McGregor and the S.M.A.R.T. lab (Mechanical and Nuclear Engineering, Kansas State University) for microwell fabrication, and Prof. Christopher Bowman (Chemical and Biological Enigneering, University of Colorado) for fruitful discussion. 1
Publisher Copyright:
Copyright © 2018 American Chemical Society.
Copyright:
Copyright 2019 Elsevier B.V., All rights reserved.
PY - 2019/1/22
Y1 - 2019/1/22
N2 - Microwell arrays are important tools for studying single cell behavior and cell-cell interactions, both in microbial and mammalian systems. However, retrieval of cells from microwell arrays with high spatial precision remains a major technical hurdle that prevents follow-up genetic and phenotypic characterization of cells within observed microwells. This work describes a new, material-based approach to grow and retrieve live bacterial cells from small (≥20 μm diameter) microwells in an array using the plant pathogen Agrobacterium tumefaciens as a model bacterium. Our approach uses a light-responsive, step-polymerized poly(ethylene glycol) hydrogel interface as a membrane that confines motile cells within microwells while allowing nutrient exchange and cell growth. The key design feature is the photodegradability of the membrane, as it enables individual wells of interest to be opened using patterned UV light for selective release and retrieval of cells. Extraction can occur in parallel from any number and combination of wells defined by the user. These advancements represent a new use for light-responsive hydrogels and the ability to retrieve cells from microwells with high spatial precision enables several applications that require the isolation and characterization of cells with rare phenotypes from heterogeneous populations.
AB - Microwell arrays are important tools for studying single cell behavior and cell-cell interactions, both in microbial and mammalian systems. However, retrieval of cells from microwell arrays with high spatial precision remains a major technical hurdle that prevents follow-up genetic and phenotypic characterization of cells within observed microwells. This work describes a new, material-based approach to grow and retrieve live bacterial cells from small (≥20 μm diameter) microwells in an array using the plant pathogen Agrobacterium tumefaciens as a model bacterium. Our approach uses a light-responsive, step-polymerized poly(ethylene glycol) hydrogel interface as a membrane that confines motile cells within microwells while allowing nutrient exchange and cell growth. The key design feature is the photodegradability of the membrane, as it enables individual wells of interest to be opened using patterned UV light for selective release and retrieval of cells. Extraction can occur in parallel from any number and combination of wells defined by the user. These advancements represent a new use for light-responsive hydrogels and the ability to retrieve cells from microwells with high spatial precision enables several applications that require the isolation and characterization of cells with rare phenotypes from heterogeneous populations.
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U2 - 10.1021/acsabm.8b00592
DO - 10.1021/acsabm.8b00592
M3 - Article
AN - SCOPUS:85064804977
VL - 2
SP - 266
EP - 276
JO - ACS Applied Bio Materials
JF - ACS Applied Bio Materials
SN - 2576-6422
IS - 1
ER -