DNA-templated assembly of droplet-derived PEG microtissues
- 21 July 2011
- journal article
- research article
- Published by Royal Society of Chemistry (RSC) in Lab on a Chip
- Vol. 11 (17), 2967-2975
- https://doi.org/10.1039/c1lc20318e
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.Keywords
This publication has 49 references indexed in Scilit:
- Hallmarks of Cancer: The Next GenerationCell, 2011
- Reconstituting Organ-Level Lung Functions on a ChipScience, 2010
- Microenvironmental regulation of the sinusoidal endothelial cell phenotype in vitroHepatology, 2009
- Human mammary progenitor cell fate decisions are products of interactions with combinatorial microenvironmentsIntegrative Biology, 2009
- Stem Cells and Niches: Mechanisms That Promote Stem Cell Maintenance throughout LifeCell, 2008
- Micromechanical control of cell–cell interactionsProceedings of the National Academy of Sciences of the United States of America, 2007
- Capturing complex 3D tissue physiology in vitroNature Reviews Molecular Cell Biology, 2006
- Microscale technologies for tissue engineering and biologyProceedings of the National Academy of Sciences of the United States of America, 2006
- Stromal fibroblasts in cancer initiation and progressionNature, 2004
- Endothelial signaling during developmentNature Medicine, 2003