Fabrication of cell-containing hydrogel microstructures inside microfluidic devices that can be used as cell-based biosensors

This paper describes microfluidic systems containing immobilized hydrogel-encapsulated mammalian cells that can be used as cell-based biosensors. Mammalian cells were encapsulated in three-dimensional poly(ethylene glycol)(PEG) hydrogel microstructures which were photolithographically polymerized in microfluidic devices and grown under static culture conditions. The encapsulated cells remained viable for a week and were able to carry out enzymatic reactions inside the microfluidic devices. Cytotoxicity assays proved that small molecular weight toxins such as sodium azide could easily diffuse into the hydrogel microstructures and kill the encapsulated cells, which resulted in decreased viability. Furthermore, heterogeneous hydrogel microstructures encapsulating two different phenotypes in discrete spatial locations were also successfully fabricated inside microchannels.

[1]  Teruo Fujii,et al.  Cell Culture in 3-Dimensional Microfluidic Structure of PDMS (polydimethylsiloxane) , 2003 .

[2]  Michael V. Pishko,et al.  Biomems Materials and Fabrication Technology: Control of Mammalian Cell and Bacteria Adhesion on Substrates Micropatterned with Poly(ethylene Glycol) Hydrogels , 2022 .

[3]  N. Peppas,et al.  Correlation between mesh size and equilibrium degree of swelling of polymeric networks. , 1989, Journal of biomedical materials research.

[4]  Aaron R Wheeler,et al.  Microfluidic device for single-cell analysis. , 2003, Analytical chemistry.

[5]  Richard M Crooks,et al.  Hydrogel-based microreactors as a functional component of microfluidic systems. , 2002, Analytical chemistry.

[6]  M. Pishko,et al.  Release of protein from highly cross-linked hydrogels of poly(ethylene glycol) diacrylate fabricated by UV polymerization. , 2001, Biomaterials.

[7]  Andreas Manz,et al.  Chip-based microsystems for genomic and proteomic analysis , 2000 .

[8]  J. Neuberger,et al.  A Bioartificial Liver--State of the Art , 2002, Science.

[9]  Alan G. R. Evans,et al.  Microfluidic Technology and Applications , 2007 .

[10]  G. Whitesides,et al.  Patterning proteins and cells using soft lithography. , 1999, Biomaterials.

[11]  T. Park,et al.  Integration of Cell Culture and Microfabrication Technology , 2003, Biotechnology progress.

[12]  S. A. Stern,et al.  Diffusion of Gases in Silicone Polymers: Molecular Dynamics Simulations , 1998 .

[13]  J. Hubbell,et al.  Characterization of permeability and network structure of interfacially photopolymerized poly(ethylene glycol) diacrylate hydrogels. , 1998, Biomaterials.

[14]  A. Mata,et al.  Characterization of Polydimethylsiloxane (PDMS) Properties for Biomedical Micro/Nanosystems , 2005, Biomedical microdevices.

[15]  Robin H. Liu,et al.  Functional hydrogel structures for autonomous flow control inside microfluidic channels , 2000, Nature.

[16]  N. Peppas Hydrogels in Medicine and Pharmacy , 1987 .

[17]  Won-Gun Koh,et al.  Poly(ethylene glycol) hydrogel microstructures encapsulating living cells. , 2002, Langmuir : the ACS journal of surfaces and colloids.

[18]  Michael V. Pishko,et al.  Photoreaction Injection Molding of Biomaterial Microstructures , 2003 .

[19]  G. Seong,et al.  Fabrication of microchambers defined by photopolymerized hydrogels and weirs within microfluidic systems: application to DNA hybridization. , 2002, Analytical chemistry.

[20]  S A Sundberg,et al.  High-throughput and ultra-high-throughput screening: solution- and cell-based approaches. , 2000, Current opinion in biotechnology.

[21]  G. Whitesides,et al.  Fabrication of microfluidic systems in poly(dimethylsiloxane) , 2000, Electrophoresis.

[22]  Milan Mrksich,et al.  Micropatterned Surfaces for Control of Cell Shape, Position, and Function , 1998, Biotechnology progress.

[23]  M L Yarmush,et al.  Controlling cell interactions by micropatterning in co-cultures: hepatocytes and 3T3 fibroblasts. , 1997, Journal of biomedical materials research.

[24]  D. Figeys,et al.  Lab-on-a-chip: a revolution in biological and medical sciences , 2000, Analytical chemistry.

[25]  R. Pal,et al.  Microfabricated reaction and separation systems. , 2001, Current opinion in biotechnology.

[26]  David J. Beebe,et al.  Insect Cell Culture in Microfluidic Channels , 2002 .

[27]  Michael V. Pishko,et al.  Amperometric Biosensors Based on Oxidoreductases Immobilized in Photopolymerized Poly(ethylene glycol) Redox Polymer Hydrogels , 1998 .

[28]  Martin A. M. Gijs,et al.  Biosystem for the culture and characterisation of epithelial cell tissues SAS , 2000 .

[29]  András Guttman,et al.  Microchip-based high-throughput screening analysis of combinatorial libraries. , 2002, Current opinion in chemical biology.

[30]  B. Ratner,et al.  In vitro study of the intrinsic toxicity of synthetic surfaces to cells. , 1994, Journal of biomedical materials research.

[31]  Jinseok Heo,et al.  A microfluidic bioreactor based on hydrogel-entrapped E. coli: cell viability, lysis, and intracellular enzyme reactions. , 2003, Analytical chemistry.

[32]  G. Whitesides,et al.  Soft lithography in biology and biochemistry. , 2001, Annual review of biomedical engineering.

[33]  Daniel I. C. Wang,et al.  Engineering cell shape and function. , 1994, Science.

[34]  Luc J. Bousse Whole cell biosensors , 1996 .

[35]  S. Bhatia,et al.  Three-Dimensional Photopatterning of Hydrogels Containing Living Cells , 2002 .

[36]  Paul C. H. Li,et al.  Transport, manipulation, and reaction of biological cells on-chip using electrokinetic effects. , 1997, Analytical chemistry.

[37]  Won-Gun Koh,et al.  Molding of hydrogel microstructures to create multiphenotype cell microarrays. , 2003, Analytical chemistry.

[38]  J. Wild,et al.  Poly(ethylene glycol) hydrogel-encapsulated fluorophore-enzyme conjugates for direct detection of organophosphorus neurotoxins. , 1999, Analytical chemistry.

[39]  L. Griffith,et al.  Tissue Engineering--Current Challenges and Expanding Opportunities , 2002, Science.

[40]  V. Yadavalli,et al.  Fabrication of poly(ethylene glycol) hydrogel microstructures using photolithography. , 2001, Langmuir : the ACS journal of surfaces and colloids.

[41]  R. Crooks,et al.  Mammalian cell cultures on micropatterned surfaces of weak-acid, polyelectrolyte hyperbranched thin films on gold. , 2001, Analytical chemistry.

[42]  G M Whitesides,et al.  Patterning cells and their environments using multiple laminar fluid flows in capillary networks. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[43]  Y. Ito,et al.  Surface micropatterning to regulate cell functions. , 1999, Biomaterials.

[44]  K. G. Olsen,et al.  Immobilization of DNA hydrogel plugs in microfluidic channels. , 2002, Analytical chemistry.

[45]  G M Whitesides,et al.  Fabrication of topologically complex three-dimensional microfluidic systems in PDMS by rapid prototyping. , 2000, Analytical chemistry.

[46]  T. Matsuda,et al.  Development of surface photochemical modification method for micropatterning of cultured cells. , 1995, Journal of biomedical materials research.

[47]  K M Shaffer,et al.  Immobilization of neural cells in three-dimensional matrices for biosensor applications. , 2000, Biosensors & bioelectronics.

[48]  M. Toner,et al.  Microengineering of cellular interactions. , 2000, Annual review of biomedical engineering.

[49]  D. L. Taylor,et al.  Topographical and Physicochemical Modification of Material Surface to Enable Patterning of Living Cells , 2001, Critical reviews in biotechnology.