Quantification of toroid microtissue contraction using a toroid climbing assay

Howes, Andrew

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Overview

Year:: 2018
Contributor:: Howes, Andrew (creator); Morgan, Jeffrey (Advisor); Mathiowitz, Edith (Reader); Achilli, Toni-Marie (Reader); Brown University. Department of Molecular Pharmacology, Physiology and Biotechnology (sponsor)
Genre:: theses
Subject:: tissue engineering
Extent:: , None p.
DOI: https://doi.org/10.26300/t0pb-qm11

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Description

Abstract:: The advancement of three dimensional biological techniques, such as microtissues, allows for the more accurate representation of in vivo structures. Yet, the added dimension pushes the limits of traditional analysis techniques. As such, there is a need for new quantitative assays for three dimensional tissues. Using non-adhesive agarose molds, microtissues such as spheroids and toroids can be created. Upon seeding into toroid shaped molds, cells self-assemble and naturally contract in a circumferential manner around a peg. These toroid microtissues can then be used as building blocks for biofabrication and or in vitro models. The toroid climbing optimization assay was developed to utilize this circumferential contraction and direct it within specifically designed non-adhesive agarose molds to quantify microtissue assembly. For a single toroid, cells were seeded into a circular trough at the base of an angled ramp surrounding a central peg. The ramp angles varied between 45 and 75 with a length of 1mm. Using wide-field microscopy, cells can be seen self-assembling into toroid microtissues and contracting up the precisely designed ramps from a bottom-view. As the inner lumen diameter changes with contraction, the position of the microtissue on the ramp can be tracked over time based on the mold geometry. This provides a more high-throughput method than the current approach of side-view based on the amount of toroids that can be tracked during a single experiment. Having a more high-throughput quantitative method provides a workable approach to conduct screening assays and proof-of-concept for the scale-up to a 96-well format.
Notes:: Thesis (Sc. M.)--Brown University, 2018

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Rights: In Copyright
Restrictions on Use: Collection is open for research.

Citation

Howes, Andrew, "Quantification of toroid microtissue contraction using a toroid climbing assay" (2018). Molecular Pharmacology, Physiology, and Biotechnology Theses and Dissertations. Brown Digital Repository. Brown University Library. https://doi.org/10.26300/t0pb-qm11

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Molecular Pharmacology, Physiology, and Biotechnology Theses and Dissertations

Theses and Dissertations for the Molecular Pharmacology, Physiology, and Biotechnology department.
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