A 3D-printed microbial cell culture platform with in situ PEGDA hydrogel barriers for differential substrate delivery

Kadilak, Andrea L.; Rehaag, Jessica C.; Harrington, Cameron A.; Shor, Leslie M.

doi:10.1063/1.5003477

Title: A 3D-printed microbial cell culture platform with in situ PEGDA hydrogel barriers for differential substrate delivery

Journal Article · Mon Oct 02 00:00:00 EDT 2017 · Biomicrofluidics

DOI:https://doi.org/10.1063/1.5003477· OSTI ID:1535346

Kadilak, Andrea L. ^[1]; Rehaag, Jessica C. ^[1]; Harrington, Cameron A. ^[1];

^[2]

Univ. of Connecticut, Storrs, CT (United States). Dept. of Chemical and Biomolecular Engineering
Univ. of Connecticut, Storrs, CT (United States). Dept. of Chemical and Biomolecular Engineering and Center for Environmental Sciences and Engineering

Additive manufacturing, or 3D-printing, techniques have recently begun to enable simpler, faster, and cheaper production of millifluidic devices at resolutions approaching 100-200 µm. At this resolution, cell culture devices can be constructed that more accurately replicate natural environments compared with conventional culturing techniques. A number of microfluidics researchers have begun incorporating additive manufacturing into their work, using 3D-printed devices in a wide array of chemical, fluidic, and even some biological applications. Here, we describe a 3D-printed cell culture platform and demonstrate its use in culturing Psedudomonas putida KT2440 bacteria for 44 h under a differential substrate gradient. PEGDA hydrogel barriers are patterned in situ within a 3D-printed channel. Transport of toluidine blue tracer dye through the hydrogel barriers is characterized. Nutrients and oxygen were delivered to cells in the culture region by diffusion through the PEGDA hydrogel barriers from adjacent media or saline perfusion channels. Expression of green fluorescent protein (GFP) by P. putida KT2440 enabled real time visualization of cell density within the 3D-printed channel, and demonstrated cells were actively expressing protein over the course of the experiment. Cells were observed clustering near hydrogel barrier boundaries where fresh substrate and oxygen were being delivered via diffusive transport, but cells were unable to penetrate the barrier. The device described here provides a versatile and easy to implement platform for cell culture in readily-controlled gradient microenvironments. By adjusting device geometry and hydrogel properties, this platform could be further customized for a wide variety of biological applications.

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Research Organization:: Univ. of Connecticut, Storrs, CT (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: SC0014522

OSTI ID:: 1535346

Alternate ID(s):: OSTI ID: 1395904; OSTI ID: 1595557

Report Number(s):: DOE-UCONN-14522-4

Journal Information:: Biomicrofluidics, Vol. 11, Issue 5; ISSN 1932-1058

Publisher:: American Institute of Physics (AIP)Copyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 11 works

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References (66)

Direct Photopolymerisation of PEG-Methacrylate Oligomers for an Easy Prototyping of Microfluidic Structures Turri, S.; Levi, M.; Emilitri, E. Macromolecular Chemistry and Physics, Vol. 211, Issue 8 https://doi.org/10.1002/macp.200900489	journal	January 2010
Diffusion: Mass Transfer in Fluid Systems Cussler, E. L. https://doi.org/10.1017/CBO9780511805134	book	January 2009
A high-throughput microfluidic assay to study neurite response to growth factor gradients Kothapalli, Chandrasekhar R.; van Veen, Ed; de Valence, Sarra Lab Chip, Vol. 11, Issue 3 https://doi.org/10.1039/C0LC00240B	journal	January 2011
Light-Activated Immobilization of Biomolecules to Agarose Hydrogels for Controlled Cellular Response Luo, Ying; Shoichet, Molly S. Biomacromolecules, Vol. 5, Issue 6 https://doi.org/10.1021/bm0495811	journal	November 2004
3D printed microfluidic devices: enablers and barriers Waheed, Sidra; Cabot, Joan M.; Macdonald, Niall P. Lab on a Chip, Vol. 16, Issue 11 https://doi.org/10.1039/C6LC00284F	journal	January 2016
The upcoming 3D-printing revolution in microfluidics Bhattacharjee, Nirveek; Urrios, Arturo; Kang, Shawn Lab on a Chip, Vol. 16, Issue 10 https://doi.org/10.1039/C6LC00163G	journal	January 2016
Cell migration into scaffolds under co-culture conditions in a microfluidic platform Chung, Seok; Sudo, Ryo; Mack, Peter J. Lab Chip, Vol. 9, Issue 2 https://doi.org/10.1039/B807585A	journal	January 2009
Development of disposable PDMS micro cell culture analog devices with photopolymerizable hydrogel encapsulating living cells Xu, Hui; Wu, Jun; Chu, Chih-Chang Biomedical Microdevices, Vol. 14, Issue 2 https://doi.org/10.1007/s10544-011-9617-4	journal	December 2011
Generation of stable concentration gradients in 2D and 3D environments using a microfluidic ladder chamber Saadi, Wajeeh; Rhee, Seog Woo; Lin, Francis Biomedical Microdevices, Vol. 9, Issue 5 https://doi.org/10.1007/s10544-007-9051-9	journal	May 2007
Synergistic effects of soil microstructure and bacterial EPS on drying rate in emulated soil micromodels Deng, Jinzi; Orner, Erika P.; Chau, Jessica Furrer Soil Biology and Biochemistry, Vol. 83 https://doi.org/10.1016/j.soilbio.2014.12.006	journal	April 2015
A platform for assessing chemotactic migration within a spatiotemporally defined 3D microenvironment Abhyankar, Vinay V.; Toepke, Michael W.; Cortesio, Christa L. Lab on a Chip, Vol. 8, Issue 9 https://doi.org/10.1039/b803533d	journal	January 2008
Diffusion-Based and Long-Range Concentration Gradients of Multiple Chemicals for Bacterial Chemotaxis Assays Kim, Minseok; Kim, Taesung Analytical Chemistry, Vol. 82, Issue 22 https://doi.org/10.1021/ac102022q	journal	November 2010
Fabrication of biocompatible lab-on-chip devices for biomedical applications by means of a 3D-printing process: Fabrication of biocompatible lab-on-chip devices for biomedical applications Takenaga, S.; Schneider, B.; Erbay, E. physica status solidi (a), Vol. 212, Issue 6 https://doi.org/10.1002/pssa.201532053	journal	May 2015
Diffusion-Based Analysis of Molecular Interactions in Microfluidic Devices Hatch, A.; Garcia, E.; Yager, P. Proceedings of the IEEE, Vol. 92, Issue 1 https://doi.org/10.1109/JPROC.2003.820547	journal	January 2004
Microfabrication meets microbiology Weibel, Douglas B.; DiLuzio, Willow R.; Whitesides, George M. Nature Reviews Microbiology, Vol. 5, Issue 3 https://doi.org/10.1038/nrmicro1616	journal	March 2007
Effects of PEG hydrogel crosslinking density on protein diffusion and encapsulated islet survival and function Weber, Laney M.; Lopez, Christina G.; Anseth, Kristi S. Journal of Biomedical Materials Research Part A, Vol. 90A, Issue 3 https://doi.org/10.1002/jbm.a.32134	journal	September 2009
Differential Effects of Permeating and Nonpermeating Solutes on the Fatty Acid Composition of Pseudomonas putida Halverson, L. J.; Firestone, M. K. Applied and Environmental Microbiology, Vol. 66, Issue 6 https://doi.org/10.1128/AEM.66.6.2414-2421.2000	journal	June 2000
Mail-order microfluidics: evaluation of stereolithography for the production of microfluidic devices Au, Anthony K.; Lee, Wonjae; Folch, Albert Lab Chip, Vol. 14, Issue 7 https://doi.org/10.1039/C3LC51360B	journal	January 2014
Three-Dimensional Patterning of Poly(Ethylene Glycol) Hydrogels Through Surface-Initiated Photopolymerization Papavasiliou, Georgia; Songprawat, Preedarat; Pérez-Luna, Victor Tissue Engineering Part C: Methods, Vol. 14, Issue 2 https://doi.org/10.1089/ten.tec.2007.0355	journal	June 2008
Characterization of the interaction between fibroblasts and tumor cells on a microfluidic co-culture device Ma, Huipeng; Liu, Tingjiao; Qin, Jianhua ELECTROPHORESIS, Vol. 31, Issue 10 https://doi.org/10.1002/elps.200900776	journal	April 2010
Assessment of the biocompatibility of three-dimensional-printed polymers using multispecies toxicity tests Zhu, Feng; Friedrich, Timo; Nugegoda, Dayanthi Biomicrofluidics, Vol. 9, Issue 6 https://doi.org/10.1063/1.4939031	journal	November 2015
Sprouting Angiogenesis under a Chemical Gradient Regulated by Interactions with an Endothelial Monolayer in a Microfluidic Platform Jeong, Gi Seok; Han, Sewoon; Shin, Yoojin Analytical Chemistry, Vol. 83, Issue 22 https://doi.org/10.1021/ac202170e	journal	November 2011
Microfluidic fabrication of microengineered hydrogels and their application in tissue engineering Chung, Bong Geun; Lee, Kwang-Ho; Khademhosseini, Ali Lab Chip, Vol. 12, Issue 1 https://doi.org/10.1039/C1LC20859D	journal	January 2012
Engineering microscale cellular niches for three-dimensional multicellular co-cultures Huang, Carlos P.; Lu, Jente; Seon, Hyeryung Lab on a Chip, Vol. 9, Issue 12 https://doi.org/10.1039/b818401a	journal	January 2009
Interplay of biomaterials and micro-scale technologies for advancing biomedical applications Khademhosseini, Ali; Bettinger, Chris; Karp, Jeffrey M. Journal of Biomaterials Science, Polymer Edition, Vol. 17, Issue 11 https://doi.org/10.1163/156856206778667488	journal	January 2006
Co-culture of epithelial cells and bacteria for investigating host–pathogen interactions Kim, Jeongyun; Hegde, Manjunath; Jayaraman, Arul Lab Chip, Vol. 10, Issue 1 https://doi.org/10.1039/B911367C	journal	January 2010
Molded polyethylene glycol microstructures for capturing cells within microfluidic channels Khademhosseini, Ali; Yeh, Judy; Jon, Sangyong Lab on a Chip, Vol. 4, Issue 5 https://doi.org/10.1039/b404842c	journal	January 2004
Let there be chip—towards rapid prototyping of microfluidic devices: one-step manufacturing processes Waldbaur, Ansgar; Rapp, Holger; Länge, Kerstin Analytical Methods, Vol. 3, Issue 12 https://doi.org/10.1039/c1ay05253e	journal	January 2011
A microfluidic device for parallel 3-D cell cultures in asymmetric environments Frisk, Thomas; Rydholm, Susanna; Liebmann, Thomas ELECTROPHORESIS, Vol. 28, Issue 24 https://doi.org/10.1002/elps.200700342	journal	December 2007
Fabrication of non-biofouling polyethylene glycol micro- and nanochannels by ultraviolet-assisted irreversible sealing Kim, Pilnam; Jeong, Hoon Eui; Khademhosseini, Ali Lab on a Chip, Vol. 6, Issue 11 https://doi.org/10.1039/b610503c	journal	January 2006
3D printed microfluidics for biological applications Ho, Chee Meng Benjamin; Ng, Sum Huan; Li, King Ho Holden Lab on a Chip, Vol. 15, Issue 18 https://doi.org/10.1039/C5LC00685F	journal	January 2015
Microfluidic device generating stable concentration gradients for long term cell culture: application to Wnt3a regulation of β-catenin signaling Cimetta, Elisa; Cannizzaro, Christopher; James, Richard Lab on a Chip, Vol. 10, Issue 23 https://doi.org/10.1039/c0lc00033g	journal	January 2010
Three-dimensional extracellular matrix-mediated neural stem cell differentiation in a microfluidic device Han, Sewoon; Yang, Kisuk; Shin, Yoojin Lab on a Chip, Vol. 12, Issue 13 https://doi.org/10.1039/c2lc21285d	journal	January 2012
3D printed microfluidic devices with integrated valves Rogers, Chad I.; Qaderi, Kamran; Woolley, Adam T. Biomicrofluidics, Vol. 9, Issue 1 https://doi.org/10.1063/1.4905840	journal	January 2015
Extracellular Polymeric Substances Affecting Pore-Scale Hydrologic Conditions for Bacterial Activity in Unsaturated Soils Or, Dani; Phutane, Sachin; Dechesne, Arnaud Vadose Zone Journal, Vol. 6, Issue 2 https://doi.org/10.2136/vzj2006.0080	journal	January 2007
Solute Diffusion within Hydrogels. Mechanisms and Models Amsden, Brian Macromolecules, Vol. 31, Issue 23 https://doi.org/10.1021/ma980765f	journal	November 1998
Configurable 3D-Printed millifluidic and microfluidic ‘lab on a chip’ reactionware devices Kitson, Philip J.; Rosnes, Mali H.; Sans, Victor Lab on a Chip, Vol. 12, Issue 18 https://doi.org/10.1039/c2lc40761b	journal	January 2012
A new method for studying gradient-induced neutrophil desensitization based on an open microfluidic chamber Keenan, Thomas M.; Frevert, Charles W.; Wu, Aileen Lab Chip, Vol. 10, Issue 1 https://doi.org/10.1039/B913494H	journal	January 2010
Covalently immobilized biomolecule gradient on hydrogel surface using a gradient generating microfluidic device for a quantitative mesenchymal stem cell study Liu, Zongbin; Xiao, Lidan; Xu, Baojian Biomicrofluidics, Vol. 6, Issue 2 https://doi.org/10.1063/1.4704522	journal	June 2012
Polymerase Chain Reaction-Capillary Electrophoresis Genetic Analysis Microdevice with In-Line Affinity Capture Sample Injection Thaitrong, Numrin; Toriello, Nicholas M.; Del Bueno, Nadia Analytical Chemistry, Vol. 81, Issue 4 https://doi.org/10.1021/ac802057f	journal	February 2009
Effect of Cross-Linking on the Diffusion of Water, Ions, and Small Molecules in Hydrogels Wu, Yanbin; Joseph, Sony; Aluru, N. R. The Journal of Physical Chemistry B, Vol. 113, Issue 11 https://doi.org/10.1021/jp808145x	journal	March 2009
Three-dimensional printed millifluidic devices for zebrafish embryo tests Zhu, Feng; Skommer, Joanna; Macdonald, Niall P. Biomicrofluidics, Vol. 9, Issue 4 https://doi.org/10.1063/1.4927379	journal	July 2015
Influence of the nature of the porous confining network on the sorption, diffusion and mechanical properties of hydrogel IPNs Ferrer, G. Gallego; Pradas, M. Monleón; Ribelles, J. L. Gómez European Polymer Journal, Vol. 46, Issue 4 https://doi.org/10.1016/j.eurpolymj.2009.12.018	journal	April 2010
Tuning the Permeability of Polymer Hydrogel Capsules: An Investigation of Cross-Linking Density, Membrane Thickness, and Cross-Linkers Chong, Siow-Feng; Lee, Ji Hyun; Zelikin, Alexander N. Langmuir, Vol. 27, Issue 5 https://doi.org/10.1021/la104510e	journal	March 2011
Cross talk between cancer and immune cells: exploring complex dynamics in a microfluidic environment Businaro, Luca; De Ninno, Adele; Schiavoni, Giovanna Lab Chip, Vol. 13, Issue 2 https://doi.org/10.1039/C2LC40887B	journal	January 2013
Remote-controlled robotic platform ORPHEUS as a new tool for detection of bacteria in the environment: Microfluidics and Miniaturization Nejdl, Lukas; Kudr, Jiri; Cihalova, Kristyna ELECTROPHORESIS, Vol. 35, Issue 16 https://doi.org/10.1002/elps.201300576	journal	April 2014
Dynamic Dosing Assay Relating Real-Time Respiration Responses of Staphylococcus aureus Biofilms to Changing Microchemical Conditions Deng, Jinzi; Dhummakupt, Adit; Samson, Philip C. Analytical Chemistry, Vol. 85, Issue 11 https://doi.org/10.1021/ac303711m	journal	May 2013
A transparent cell-culture microchamber with a variably controlled concentration gradient generator and flow field rectifier Cheng, Ji-Yen; Yen, Meng-Hua; Kuo, Ching-Te Biomicrofluidics, Vol. 2, Issue 2 https://doi.org/10.1063/1.2952290	journal	June 2008
Release of protein from highly cross-linked hydrogels of poly(ethylene glycol) diacrylate fabricated by UV polymerization Mellott, Michael B.; Searcy, Katherine; Pishko, Michael V. Biomaterials, Vol. 22, Issue 9 https://doi.org/10.1016/S0142-9612(00)00258-1	journal	May 2001
Assessment of biocompatibility of 3D printed photopolymers using zebrafish embryo toxicity assays Macdonald, N. P.; Zhu, F.; Hall, C. J. Lab on a Chip, Vol. 16, Issue 2 https://doi.org/10.1039/C5LC01374G	journal	January 2016
Assessing and Reducing the Toxicity of 3D-Printed Parts Oskui, Shirin Mesbah; Diamante, Graciel; Liao, Chunyang Environmental Science & Technology Letters, Vol. 3, Issue 1 https://doi.org/10.1021/acs.estlett.5b00249	journal	November 2015
A 3D Printed Fluidic Device that Enables Integrated Features Anderson, Kari B.; Lockwood, Sarah Y.; Martin, R. Scott Analytical Chemistry, Vol. 85, Issue 12 https://doi.org/10.1021/ac4009594	journal	May 2013
3D printed auto-mixing chip enables rapid smartphone diagnosis of anemia Plevniak, Kimberly; Campbell, Matthew; Myers, Timothy Biomicrofluidics, Vol. 10, Issue 5 https://doi.org/10.1063/1.4964499	journal	September 2016
Dimethacrylate network formation and polymer property evolution as determined by the selection of monomers and curing conditions Stansbury, Jeffrey W. Dental Materials, Vol. 28, Issue 1 https://doi.org/10.1016/j.dental.2011.09.005	journal	January 2012
Fabrication of Gradient Hydrogels Using a Microfluidics/Photopolymerization Process Burdick, Jason A.; Khademhosseini, Ali; Langer, Robert Langmuir, Vol. 20, Issue 13 https://doi.org/10.1021/la049298n	journal	June 2004
Protist-Facilitated Particle Transport Using Emulated Soil Micromodels Rubinstein, Rebecca L.; Kadilak, Andrea L.; Cousens, Virginia C. Environmental Science & Technology, Vol. 49, Issue 3 https://doi.org/10.1021/es503424z	journal	January 2015
A small-scale air-cathode microbial fuel cell for on-line monitoring of water quality Di Lorenzo, Mirella; Thomson, Alexander R.; Schneider, Kenneth Biosensors and Bioelectronics, Vol. 62 https://doi.org/10.1016/j.bios.2014.06.050	journal	December 2014
Complete genome sequence and comparative analysis of the metabolically versatile Pseudomonas putida KT2440 Nelson, K. E.; Weinel, C.; Paulsen, I. T. Environmental Microbiology, Vol. 4, Issue 12, p. 799-808 https://doi.org/10.1046/j.1462-2920.2002.00366.x	journal	December 2002
A ubiquitin-based tagging system for controlled modulation of protein stability Stack, Jeffrey H.; Whitney, Michael; Rodems, Steven M. Nature Biotechnology, Vol. 18, Issue 12 https://doi.org/10.1038/82422	journal	December 2000
3D-Printed Microfluidic Device for the Detection of Pathogenic Bacteria Using Size-based Separation in Helical Channel with Trapezoid Cross-Section Lee, Wonjae; Kwon, Donghoon; Choi, Woong Scientific Reports, Vol. 5, Issue 1 https://doi.org/10.1038/srep07717	journal	January 2015
Design and additive manufacture for flow chemistry Capel, Andrew J.; Edmondson, Steve; Christie, Steven D. R. Lab on a Chip, Vol. 13, Issue 23 https://doi.org/10.1039/c3lc50844g	journal	January 2013
Laminar-Turbulent Transition in Ducts of Rectangular Cross Section Hanks, R. W.; Ruo, H. -C. Industrial & Engineering Chemistry Fundamentals, Vol. 5, Issue 4 https://doi.org/10.1021/i160020a022	journal	November 1966
Advances in three-dimensional rapid prototyping of microfluidic devices for biological applications O'Neill, P. F.; Ben Azouz, A.; Vázquez, M. Biomicrofluidics, Vol. 8, Issue 5 https://doi.org/10.1063/1.4898632	journal	September 2014
3D-printed microfluidic automation Au, Anthony K.; Bhattacharjee, Nirveek; Horowitz, Lisa F. Lab on a Chip, Vol. 15, Issue 8 https://doi.org/10.1039/C5LC00126A	journal	January 2015
Drug Release from Interpenetrating Polymer Networks Based on Poly(ethylene glycol) Methyl Ether Acrylate and Gelatin Ding, Frank; Hsu, S. -H.; Wu, D. -H. Journal of Biomaterials Science, Polymer Edition, Vol. 20, Issue 5-6 https://doi.org/10.1163/156856209X426420	journal	January 2009
Complete genome sequence and comparative analysis of the metabolically versatile Pseudomonas putida KT2440 Nelson, K. E.; Weinel, C.; Paulsen, I. T. Environmental Microbiology, Vol. 5, Issue 7 https://doi.org/10.1046/j.1462-2920.2003.00480.x	journal	July 2003

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