DNA-templated assembly of droplet-derived PEG microtissues
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...
| Main Authors: | , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | en_US |
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Royal Society of Chemistry, The
2012
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| Online Access: | http://hdl.handle.net/1721.1/75312 https://orcid.org/0000-0002-1293-2097 |
| _version_ | 1826215354387398656 |
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| author | Li, Cheri Yingjie Wood, David K. Hsu, Caroline M. Bhatia, Sangeeta N. |
| author2 | Harvard University--MIT Division of Health Sciences and Technology |
| author_facet | Harvard University--MIT Division of Health Sciences and Technology Li, Cheri Yingjie Wood, David K. Hsu, Caroline M. Bhatia, Sangeeta N. |
| author_sort | Li, Cheri Yingjie |
| collection | MIT |
| description | 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 (10[superscript 5] h[superscript −1]). Multiple cell types, including suspension and adherently cultured cells, can be encapsulated into the microtissues with high viability ([similar]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. |
| first_indexed | 2024-09-23T16:26:18Z |
| format | Article |
| id | mit-1721.1/75312 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T16:26:18Z |
| publishDate | 2012 |
| publisher | Royal Society of Chemistry, The |
| record_format | dspace |
| spelling | mit-1721.1/753122022-10-02T07:57:48Z DNA-templated assembly of droplet-derived PEG microtissues Li, Cheri Yingjie Wood, David K. Hsu, Caroline M. Bhatia, Sangeeta N. Harvard University--MIT Division of Health Sciences and Technology Massachusetts Institute of Technology. Department of Chemical Engineering Massachusetts Institute of Technology. Department of Chemical Engineering Li, Cheri Yingjie Wood, David K. Hsu, Caroline M. Bhatia, Sangeeta N. 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 (10[superscript 5] h[superscript −1]). Multiple cell types, including suspension and adherently cultured cells, can be encapsulated into the microtissues with high viability ([similar]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. 2012-12-10T15:08:00Z 2012-12-10T15:08:00Z 2011-07 2011-04 Article http://purl.org/eprint/type/JournalArticle 1473-0197 1473-0189 http://hdl.handle.net/1721.1/75312 Li, Cheri Y. et al. “DNA-templated Assembly of Droplet-derived PEG Microtissues.” Lab on a Chip 11.17 (2011): 2967. https://orcid.org/0000-0002-1293-2097 en_US http://dx.doi.org/10.1039/c1lc20318e Lab on a Chip Creative Commons Attribution-Noncommercial-Share Alike 3.0 http://creativecommons.org/licenses/by-nc-sa/3.0/ application/pdf Royal Society of Chemistry, The PMC |
| spellingShingle | Li, Cheri Yingjie Wood, David K. Hsu, Caroline M. Bhatia, Sangeeta N. DNA-templated assembly of droplet-derived PEG microtissues |
| title | DNA-templated assembly of droplet-derived PEG microtissues |
| title_full | DNA-templated assembly of droplet-derived PEG microtissues |
| title_fullStr | DNA-templated assembly of droplet-derived PEG microtissues |
| title_full_unstemmed | DNA-templated assembly of droplet-derived PEG microtissues |
| title_short | DNA-templated assembly of droplet-derived PEG microtissues |
| title_sort | dna templated assembly of droplet derived peg microtissues |
| url | http://hdl.handle.net/1721.1/75312 https://orcid.org/0000-0002-1293-2097 |
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