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Issue 17, 2011
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DNA-templated assembly of droplet-derived PEG microtissues

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Abstract

Patterning multiple cell types is a critical step for engineering functional tissues, but few methods provide three-dimensional positioning at the cellular length scale. Here, we present a “bottom-up” approach for fabricating multicellular tissue constructs that utilizes DNA-templated assembly of 3D cell-laden hydrogel microtissues. A flow focusing-generated emulsion of photopolymerizable prepolymer is used to produce 100 μm monodisperse microtissues at a rate of 100 Hz (105 h−1). Multiple cell types, including suspension and adherently cultured cells, can be encapsulated into the microtissues with high viability (∼97%). We then use a DNA coding scheme to self-assemble microtissues “bottom-up” from a template that is defined using “top-down” techniques. The microtissues are derivatized with single-stranded DNA using a biotin–streptavidin linkage to the polymer network, and are assembled by sequence-specific hybridization onto spotted DNA microarrays. Using orthogonal DNA codes, we achieve multiplexed patterning of multiple microtissue types with high binding efficiency and >90% patterning specificity. Finally, we demonstrate the ability to organize multicomponent constructs composed of epithelial and mesenchymal microtissues while preserving each cell type in a 3D microenvironment. The combination of high throughput microtissue generation with scalable surface-templated assembly offers the potential to dissect mechanisms of cell–cell interaction in three dimensions in healthy and diseased states, as well as provides a framework for templated assembly of larger structures for implantation.

Graphical abstract: DNA-templated assembly of droplet-derived PEG microtissues

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Publication details

The article was received on 14 Apr 2011, accepted on 15 Jun 2011 and first published on 21 Jul 2011


Article type: Paper
DOI: 10.1039/C1LC20318E
Citation: Lab Chip, 2011,11, 2967-2975
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    DNA-templated assembly of droplet-derived PEG microtissues

    C. Y. Li, D. K. Wood, C. M. Hsu and S. N. Bhatia, Lab Chip, 2011, 11, 2967
    DOI: 10.1039/C1LC20318E

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