A 3D Bioprinter Specifically Designed for the High-Throughput Production of Matrix-Embedded Multicellular Spheroids

Robert H Utama; Lakmali Atapattu; Aidan P O'Mahony; Christopher M Fife; Jongho Baek; Théophile Allard; Kieran J O'Mahony; Julio C C Ribeiro; Katharina Gaus; Maria Kavallaris; J Justin Gooding

doi:10.1016/j.isci.2020.101621

A 3D Bioprinter Specifically Designed for the High-Throughput Production of Matrix-Embedded Multicellular Spheroids

iScience. 2020 Sep 28;23(10):101621. doi: 10.1016/j.isci.2020.101621. eCollection 2020 Oct 23.

Authors

Robert H Utama^{1

2}, Lakmali Atapattu^{1

3}, Aidan P O'Mahony⁴, Christopher M Fife^{1

3}, Jongho Baek^{5

6}, Théophile Allard⁴, Kieran J O'Mahony⁷, Julio C C Ribeiro⁴, Katharina Gaus^{5

6}, Maria Kavallaris^{1

3}, J Justin Gooding^{1

2}

Affiliations

¹ ARC Centre of Excellence in Convergent Bio-Nano Science and Technology and Australian Centre for NanoMedicine, The University of New South Wales, Sydney, NSW 2052, Australia.
² School of Chemistry, The University of New South Wales, Sydney, NSW 2052, Australia.
³ Children's Cancer Institute, Lowy Cancer Research Centre, The University of New South Wales, Sydney, NSW 2052, Australia.
⁴ Inventia Life Science Pty Ltd, Sydney, NSW, Australia.
⁵ EMBL Australia Node in Single Molecule Science, School of Medical Sciences, The University of New South Wales, Sydney, NSW 2052, Australia.
⁶ ARC Centre of Excellence in Advanced Molecular Imaging, The University of New South Wales, Sydney, NSW 2052, Australia.
⁷ OMKO Limited, Tooreen South, Bantry, Co. Cork, Ireland.

Abstract

3D in vitro cancer models are important therapeutic and biological discovery tools, yet formation of matrix-embedded multicellular spheroids prepared in high-throughput (HTP), and in a highly controlled manner, remains challenging. This is important to achieve robust and statistically relevant data. Here, we developed an enabling technology consisting of a bespoke drop-on-demand 3D bioprinter capable of HTP printing of 96-well plates of spheroids. 3D multicellular spheroids are embedded inside a hydrogel matrix with precise control over size and cell number, with the intra-experiment variability of embedded spheroid diameter coefficient of variation being between 4.2% and 8.7%. Application of 3D bioprinting HTP drug screening was demonstrated with doxorubicin. Measurements of IC₅₀ values showed sensitivity to spheroid size, embedding, and how spheroids conform to the embedding, revealing parameters shaping biological responses in these models. Our study demonstrates the potential of 3D bioprinting as a robust HTP platform to screen biological and therapeutic parameters.

Keywords: Biomaterials; Biotechnology; Cell Biology.