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Localized induction of gene expression in embryonic stem cell aggregates using holographic optical tweezers to create biochemical gradients

Kirkham, Glen R.; Ware, James; Upton, Thomas; Allen, Stephanie; Shakesheff, Kevin M.; Buttery, Lee D.K.

Authors

Glen R. Kirkham

James Ware

Thomas Upton

Stephanie Allen

Lee D.K. Buttery



Abstract

Three dimensional (3D) cell models that mimic the structure and function of native tissues are enabling more detailed study of physiological and pathological mechanisms in vitro. We have previously demonstrated the ability to build and manipulate 3D multicellular microscopic structures using holographic optical tweezers (HOTs). Here we show the construction of a precisely patterned 3D microenvironment and biochemical gradient model consisting of mouse embryoid bodies (mEBs) and polymer microparticles loaded with retinoic acid (RA), embedded in a hydrogel. We demonstrate discrete, zonal expression of the RA-inducible protein Stra8 within mEBs in response to release of RA from polymer microparticles, corresponding directly to the defined 3D positioning of the microparticles using HOTs. These results demonstrate the ability of this technology to create chemical micro-gradients at definable length scales and to elicit, with fidelity and precision, specific biological responses. This technique can be used in the study of in vitro microenvironments to enable new insights on 3D cell models, their cellular assembly and the delivery of drug or biochemical molecules for engineering and interrogation of functional and morphogenic responses.

Journal Article Type Article
Publication Date Aug 26, 2019
Print ISSN 2364-4133
Publisher Springer
Peer Reviewed Peer Reviewed
APA6 Citation Kirkham, G. R., Ware, J., Upton, T., Allen, S., Shakesheff, K. M., & Buttery, L. D. (2019). Localized induction of gene expression in embryonic stem cell aggregates using holographic optical tweezers to create biochemical gradients. Regenerative Engineering and Translational Medicine, doi:10.1007/s40883-019-00114-5
DOI https://doi.org/10.1007/s40883-019-00114-5
Publisher URL https://link.springer.com/article/10.1007%2Fs40883-019-00114-5

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