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Nanolitre liquid patterning in aqueous environments for spatially defined reagent delivery to mammalian cells

Nature Materials volume 8, pages 736–741 (2009)Cite this article

Abstract

Microscale biopatterning enables regulation of cell–material interactions1,2 and cell shape3, and enables multiplexed high-throughput studies4,5,6,7,8 in a cell- and reagent-efficient manner. The majority of available techniques rely on physical contact of a stamp3, pin8, or mask9,10 with mainly a dry surface. Inkjet and piezoelectric printing11 is carried out in a non-contact manner but still requires a substantially dry substrate to ensure fidelity of printed patterns. These existing methods, therefore, are limited for patterning onto delicate surfaces of living cells because physical contact or substantially dry conditions are damaging to them. Microfluidic patterning with laminar streams12,13 does enable non-contact patterning in fully aqueous environments but with limited throughput and reagent diffusion across interfacial flows. Here, we describe a polymeric aqueous two-phase system that enables patterning nanolitres of a reagent-containing aqueous phase, in arbitrary shapes, within a second aqueous phase covering a cell monolayer. With the appropriate medium formulation, reagents of interest remain confined to the patterned phase without significant diffusion. The fully aqueous environment ensures high reagent activity and cell viability. The utility of this strategy is demonstrated with patterned delivery of genetic materials to mammalian cells for phenotypic screening of gene expression and gene silencing.

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Figure 1: Polymeric aqueous two-phase systems generate user-defined patterns of a reagent on a cell monolayer.
Figure 2: Addressable delivery of nucleic acids to cells using two-phase patterned microarrays.
Figure 3: Patterned microarrays of lentiviral-mediated gene expression and gene knockdown.
Figure 4: Patterned microarrays facilitate phenotypic screening of function of different genes in cell cultures on soft substrates.

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Acknowledgements

The financial support for this research was provided by a gift from J. Passino and NIH grants P50CA093990 and R01CA136553. H.T. acknowledges a postdoctoral fellowship (NSERC PDF-329449-2006). We thank M. El-Sayed and Y.-L. Lin for spectrophotometry measurements.

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Authors and Affiliations

  1. Department of Biomedical Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA

    H. Tavana, A. Jovic, B. Mosadegh, Q. Y. Lee & S. Takayama

  2. Division of Molecular Medicine and Genetics, Department of Internal Medicine, The Life Sciences Institute, University of Michigan, Ann Arbor, Michigan 48109, USA

    X. Liu & S. J. Weiss

  3. Department of Radiology, University of Michigan, Ann Arbor, Michigan 48109, USA

    K. E. Luker & G. D. Luker

  4. Department of Microbiology and Immunology, University of Michigan, Ann Arbor, Michigan 48109, USA

    G. D. Luker

  5. Macromolecular Science & Engineering Center, University of Michigan, Ann Arbor, Michigan 48109, USA

    S. Takayama

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  1. H. Tavana
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Contributions

H.T. designed and carried out experiments, analysed the data and wrote the manuscript. A.J. helped optimize transfection experiments. B.M. helped with patterning experiments and imaging microarrays. Q.Y.L. carried out image processing of transfected cell microarrays. X.L. prepared collagen and helped with the collagen degradation assay. K.E.L. prepared lentiviruses. G.D.L. helped design viral transduction experiments and optimize them. S.J.W. helped design the collagen degradation assay. S.T. designed the project and edited the manuscript. All authors commented on the manuscript.

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Correspondence to S. Takayama.

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Tavana, H., Jovic, A., Mosadegh, B. et al. Nanolitre liquid patterning in aqueous environments for spatially defined reagent delivery to mammalian cells. Nature Mater 8, 736–741 (2009). https://doi.org/10.1038/nmat2515

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