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Tumor Microenvironment pp 167–196Cite as

Methods: Using Three-Dimensional Culture (Spheroids) as an In Vitro Model of Tumour Hypoxia

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Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 899))

Abstract

Regions of hypoxia in tumours can be modelled in vitro in 2D cell cultures with a hypoxic chamber or incubator in which oxygen levels can be regulated. Although this system is useful in many respects, it disregards the additional physiological gradients of the hypoxic microenvironment, which result in reduced nutrients and more acidic pH. Another approach to hypoxia modelling is to use three-dimensional spheroid cultures. In spheroids, the physiological gradients of the hypoxic tumour microenvironment can be inexpensively modelled and explored. In addition, spheroids offer the advantage of more representative modelling of tumour therapy responses compared with 2D culture. Here, we review the use of spheroids in hypoxia tumour biology research and highlight the different methodologies for spheroid formation and how to obtain uniformity. We explore the challenge of spheroid analyses and how to determine the effect on the hypoxic versus normoxic components of spheroids. We discuss the use of high-throughput analyses in hypoxia screening of spheroids. Furthermore, we examine the use of mathematical modelling of spheroids to understand more fully the hypoxic tumour microenvironment.

*These authors contributed equally.

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Author notes

    Authors and Affiliations

    1. Nuffield Division of Clinical Laboratory Sciences, Department of Oncology, University of Oxford, Oxford, OX3 9DU, UK

      Russell Leek

    2. Gray Laboratory, Cancer Research UK/MRC Oxford Institute for Radiation Oncology, University of Oxford, Old Road Campus Research Building, off Roosevelt Drive, Oxford, OX3 7DQ, UK

      David Robert Grimes

    3. Department of Oncology, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, OX3 9DS, UK

      Adrian L. Harris

    4. Cancer Biology, Division of Cancer and Stem Cells, University of Nottingham, QMC, D Floor, West Block, W/D/1374, Nottingham, NG7 2UH, UK

      Alan McIntyre

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    1. Russell Leek
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    Correspondence to Alan McIntyre .

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    Editors and Affiliations

    1. Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA

      Constantinos Koumenis

    2. Cell & Developmental Biology, Oregon Health & Science University, Portland, Oregon, USA

      Lisa M. Coussens

    3. Department of Radiation Oncology, Stanford University School of Medic, Stanford, California, USA

      Amato Giaccia

    4. Department of Oncology, University of Oxford, Oxford, Oxfordshire, United Kingdom

      Ester Hammond

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    Leek, R., Grimes, D.R., Harris, A.L., McIntyre, A. (2016). Methods: Using Three-Dimensional Culture (Spheroids) as an In Vitro Model of Tumour Hypoxia. In: Koumenis, C., Coussens, L., Giaccia, A., Hammond, E. (eds) Tumor Microenvironment. Advances in Experimental Medicine and Biology, vol 899. Springer, Cham. https://doi.org/10.1007/978-3-319-26666-4_10

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