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. 2021 Jan 28;8:623889. doi: 10.3389/fcell.2020.623889

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Copyright © 2021 Ntamo, Samodien, Burger, Muller, Muller and Chellan.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Viability of 2D monolayers and 3D spheroids. The ATP luminescence was assessed in the 2D monolayers and 3D spheroids after 24 h and 1 day in culture, respectively. ATP/mg was higher in the 3D spheroids compared to 2D monolayers (A). In the 3D spheroids, ATP content increased from day 1 to 30 (B), while glucose utilization was comparable throughout (C). The sphere surface area of the spheroids increased from day 1 to 10 and stabilized from day 14 to 19, followed by a gradual increase up to day 30 (D,E). Scale bars: 200 μm (D). Data expressed as mean ± SEM, Student’s t-test and ANOVA was used to conduct analysis: ^#p < 0.05 2D vs. 3D condition (Two-way ANOVA); **p < 0.001 vs. day 1, ***p < 0.0001 vs. day 1 (Ordinary One-way ANOVA).