Multiplexing Spheroid Volume, Resazurin and Acid Phosphatase Viability Assays for High-Throughput Screening of Tumour Spheroids and Stem Cell Neurospheres

  • Delyan P. Ivanov, Terry L. Parker, David A. Walker, Cameron Alexander, Marianne B. Ashford, Paul R. Gellert, Martin C. Garnett
  • PLoS ONE, August 2014, Public Library of Science (PLoS)
  • DOI: 10.1371/journal.pone.0103817

Spheroid health using volume, metabolism and enzymatic activity

What is it about?

Finding new cures for cancer depends on our ability to faithfully reproduce cancer biology in the lab. Patient tumours resemble small balls and are nothing like the conventional carpet-like monolayers on Petri dishes used in most labs. Mainstream adoption of physiological 3D models has been stalled in the past 50 years due to high price, low speed and poor reproducibility. This study describes a readily available, low-cost, open-source platform for spheroid cancer screens that any lab can use.

Why is it important?

Spheroids are famous for their relevance to patient disease, but notorious for reproducibility and cost. Here we show how to culture spheroids the easy way, without the need for specialized equipment and how to multiplex different methods for 3D culture health assessment. Apart from lots of useful technical tips the paper also includes an open-source image analysis macro to aid in spheroid size automation. Before you start working with spheroids read this article it will save you time and money.


Dr Delyan Pavlov Ivanov
University of Nottingham

When I first tried to culture spheroids, I found it frustratingly hard. The methods either required fancy, expensive plates, were cumbersome and slow or yielded spheroids of all sizes. Furthermore, there was very little information on the suitability of different assays for spheroid health assessment. This paper aims to show one of the easiest and most reliable ways to culture spheroids in the lab. It highlights the steps necessary to validate that your assay gives you a reliable estimate of spheroid viability.

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The following have contributed to this page: Dr Delyan Pavlov Ivanov