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. 2020 Oct 28;8:594090. doi: 10.3389/fcell.2020.594090

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Copyright © 2020 Ha, Kang, Lee, Baek, Kim, Chung, Lee and Kim.

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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Comparison of neuronal differentiation potential in 2D vs. 3D. (A,B) Immunofluorescence analysis to compare neuronal differentiation potential between 2D and 3D with neuronal markers (TUJ1, MAP2, and, NEUN) in simBOs at day 10 (A) and 20 (B) of spontaneous differentiation. Scale bars represent 100 μm. (C) Quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis for neural progenitor marker (PAX6), pan-neuronal markers (DCX, MAP2, NEUN, and SYN1), glutamatergic neuronal marker (GRIN1), GABAergic neuronal marker (GRIN2B) and glial marker (S100B) in simplified brain organoids (simBOs; 3D) and 2D differentiation samples. Expression value was normalized to those of human primitive neural stem cells (pNSC) in 2D (d0). Data represent mean ± standard error (SE) from three technical replicates. Statistical significance of Sidak’s multiple comparisons *p < 0.05, **p < 0.01, ***p < 0.001 after two-way ANOVA.