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

Kirkham, G.R. ORCID: 0000-0001-6477-2833, Ware, J., Upton, T., Allen, S., Shakesheff, K.M. and Buttery, L.D.K., 2019. Localized induction of gene expression in embryonic stem cell aggregates using holographic optical tweezers to create biochemical gradients. Regenerative Engineering and Translational Medicine. ISSN 2364-4133

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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 microgradients 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.

Item Type: Journal article
Publication Title: Regenerative Engineering and Translational Medicine
Creators: Kirkham, G.R., Ware, J., Upton, T., Allen, S., Shakesheff, K.M. and Buttery, L.D.K.
Publisher: Springer
Date: 26 August 2019
ISSN: 2364-4133
Identifiers:
NumberType
10.1007/s40883-019-00114-5DOI
Rights: © the author(s) 2019. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
Divisions: Schools > School of Science and Technology
Depositing User: Jonathan Gallacher
Date Added: 30 Sep 2019 13:39
Last Modified: 30 Sep 2019 13:41
URI: http://irep.ntu.ac.uk/id/eprint/37880

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