Reprinted from: Publication of this reprint collection is supported by paid advertising SLAS Technology 27 (2022) 267–275 Contents lists available at ScienceDirect SLAS Technology journal homepage: www.elsevier.com/locate/slast A Biomimetic High Throughput Model of Cancer Cell Spheroid Dissemination onto Aligned Fibrillar Collagen † Hossam Ibrahim1,2 , Stephen D. Thorpe 2,3 , Michael Paukshto 4 , Tatiana S. Zaitseva 4 , Wolfgang Moritz 5 , Brian J. Rodriguez 1,2,∗ 1School of Physics, University College Dublin, Belfield, Dublin 4, D04 V1W8, Ireland 2Conway Institute of Biomolecular & Biomedical Research, University College Dublin, Belfield, Dublin 4, D04 V1W8, Ireland 3UCD School of Medicine, University College Dublin, Belfield, Dublin 4, D04 V1W8, Ireland 4Fibralign Corporation, 32930 Alvarado-Niles Rd Suite 350, Union City, CA 94587, USA 5InSphero AG, Wagistrasse 27, 8952 Schlieren, Switzerland a r t i c l e i n f o Keywords: spheroid culture collagen cell migration image analysis in vitro models a b s t r a c t Cell dissemination during tumor development is a characteristic of cancer metastasis. Dissemination from threedimensional spheroid models on extracellular matrices designed to mimic tissue-specific physiological microenvironments may allow us to better elucidate the mechanism behind cancer metastasis and the response to therapeutic agents. The orientation of fibrillar collagen plays a key role in cellular processes and mediates metastasis through contact-guidance. Understanding how cells migrate on aligned collagen fibrils requires in vitro assays with reproducible and standardized orientation of collagen fibrils on the macro-to-nanoscale. Herein, we implement a spheroid-based migration assay, integrated with a fibrillar type I collagen matrix, in a manner compatible with high throughput image acquisition and quantitative analysis. The migration of highly proliferating U2OS osteosarcoma cell spheroids onto an aligned fibrillar type I collagen matrix was quantified. Cell dissemination from the spheroid was polarized with increased invasion in the direction of fibril alignment. The resulting area of cell dissemination had an aspect ratio of 1.2 ± 0.1 and an angle of maximum invasion distance of 5° ± 44° relative to the direction of collagen fibril alignment. The assay described here can be applied to a fully automated imaging and analysis pipeline for the assessment of tumor cell migration with high throughput screening. Introduction Cell migration is a critical cell function that dictates the position of cells within the body and plays a key role in embryonic development, wound healing, and metastasis. Cells can migrate by means of a broad repertoire of processes which can be described as single-cell (e.g., amoeboid, mesenchymal) and collective migration (cell sheets, clusters, and streams). [1] Commonly-employed two-dimensional (2D) Boyden chamber and scratch wound assays utilize flat monocultures that do not account for heterogeneity in nutrient and oxygen gradients or cell-cell interactions. [2,3] The former is based on two medium-containing chambers separated by a rigid porous membrane through which cells transmigrate, often in response to a chemokine gradient, with small pores impeding collective migration. [4] This method is not optimal for realtime imaging as cells can become trapped within the porous membrane. † This publication has emanated from research supported in part by a grant from Science Foundation Ireland under Grant number SFI/17/CDA/4637. For the purpose of open access, the authors have applied a CC BY public copyright license to any Author Accepted Manuscript version arising from this submission. ∗ Corresponding Author. Brian J. Rodriguez, School of Physics and Conway Institute of Biomolecular & Biomedical Research, University College Dublin, Belfield, Dublin, D04 V1W8, Ireland E-mail address: brian.rodriguez@ucd.ie (B.J. Rodriguez). The scratch wound assay is typically performed on a rigid 2D tissue culture plate, which can be coated with extracellular matrix (ECM) and involves the migration of cells from dense monolayer sheets to occupy empty regions. [4] Planar 2D monoculture models have been used previously to recapitulate the mechanical basis of wound closure dynamics of epithelial sheets, [5,6] but they tend to fail in describing migration in fibrous tissue. Cells cultured on patterned and grooved interfaces change their shape, orientation, and direction of movement in response to topographical cues through a phenomenon called contact-guidance. Contactguidance enables cells to take the path of least resistance when confronted with heterogeneous environments, supporting the notion of preferential migration along ECM interfaces, or aligned structures. Current advances in micro-nanotechnologies and biomaterial engineering offer high-level precision and repeatability in manufacturing structured temhttps://doi.org/10.1016/j.slast.2022.05.001 Received 15 February 2022; Received in revised form 26 April 2022; Accepted 12 May 2022 2472-6303/© 2022 The Authors. Published by Elsevier Inc. on behalf of Society for Laboratory Automation and Screening. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)
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