Assay and Drug Development Technologies - 17

Drug Discovery Approaches Utilizing
Three-Dimensional Cell Culture
Sarah-Louise Ryan,1,2 Anne-Marie Baird,1-3 Gisela Vaz,4
Aaron J. Urquhart,1,2 Mathias Senge,4 Derek J. Richard,1
Kenneth J. O'Byrne,1,3,5 and Anthony M. Davies2,6
1Cancer and Ageing Research Program, Institute ofHealth
and Biomedical Innovation, Queensland University of
Technology, Brisbane, Australia.
2Translational Cell Imaging Queensland, Institute ofHealth
and Biomedical Innovation, Queensland University
ofTechnology, Brisbane, Australia.
3Thoracic Oncology Research Group, Institute ofMolecular
Medicine, Trinity College Dublin, Dublin, Ireland.
4Medical Chemistry Research Group, Institute ofMolecular
Medicine, Trinity College Dublin, Dublin, Ireland.
5Division ofCancer Services, Princess Alexandra Hospital,
Brisbane, Australia.
6Irish National Centre for High Content Screening and Analysis,
Institute ofMolecular Medicine, Trinity College Dublin,
Dublin, Ireland.
ABSTRACT
Historically, two-dimensional (2D) cell culture has been the
preferred method ofproducing disease models in vitro. Recently,
there has been a move away from 2D culture in favor of generating
three-dimensional (3D) multicellular structures, which
are thought to be more representative of the in vivo environment.
This transition has brought with it an influx of technologies
capable of producing these structures in various ways.
However, it is becoming evident that many ofthese technologies
do not perform well in automated in vitro drug discovery units.
We believe that this is a result oftheir incompatibility with highthroughput
screening (HTS). In this study, we review a number of
technologies, which are currently available for producing in vitro
3D disease models. We assess their amenability with highcontent
screening and HTS and highlight our own work in attempting
to address many of the practical problems that are
hampering the successful deployment of 3D cell systems in
mainstream research.
INTRODUCTION
O
ver the last century, there has been a steady
progression in the diversity and complexity of
tissue culture methodologies used in biomedical
research, the earliest reported examples of in vitro
cell culture being that conducted by Ross G. Harrison who
observed neuronal sprouting from frog embryo spinal cords on
a microscope slide in 1907.1 At present, cell-based research is
often performed on a variety of planar surfaces that have been
modified to promote the growth of two-dimensional (2D) cellular
monolayers. These monolayers are utilized for majority of
in vitro evaluations in research and have proven very effective.
However, it is evident that while these approaches provide a
convenient means of treating and analyzing cells, they do not
reliably permit the formation ofmulticellular structures, which
in turn form microenvironments similar to that found in vivo.2-
8 Hence the interest in generating more biologically relevant
in vitromodels, such as three-dimensional (3D) culture systems.
Limitations associated with 2D models have been identified;
such as the loss of tissue-specific architecture, mechanical
and biochemical cues, and cell-to-cell interactions.9-12 Conversely,
the microenvironment generated by 3D cell culture
appears more representative ofthat observed in vivo, resulting
in relevant cell-to-cell and cell-to-extracellular matrix
(ECM) signalling.13-16 Such signalling cascades are deemed
essential for a multitude of cellular processes, including
differentiation and proliferation.9,17,18
In contrast to conventional 2D methods, cells cultured in a 3D
format may exhibit unique biochemical and morphological
features similar to their corresponding tissues in vivo19 (summarized
in Table 1). It should be noted that the cell type, aswell as
the 3D culture method, impacts on cell organization and formation
of the 3D structure. However, the concentric arrangement
ofheterogeneous cell populations in 3D cultures, as well as
their growth pattern, mimics the initial (i) avascular stages of
solid tumors in vivo, (ii) not-yet-vascularized micrometastatic
foci, (iii) intercapillary tumor microregions with a high
proliferative activity close to the capillaries, (iv) quiescent cells as
intermediates, and (v) necrotic areas at larger distances.20,21
Two-dimensional cell-based assays are well established in
Reprinted with permission from Assay Drug Dev Technol 2016;14(1):19-28.
the drug discovery process, particularly in cancer.22 However,
their value in predicting clinical responses to new agents is
DOI: 10.1089/adt.2015.670
ª 2022 MARY ANN LIEBERT, INC. ASSAY and Drug Development Technologies 17
Assay and Drug Development Technologies

Table of Contents for the Digital Edition of Assay and Drug Development Technologies
