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. 2021 Jun 3;11:11695. doi: 10.1038/s41598-021-91203-5

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Axon extension of iPSC motor neuron spheroids is influenced by developmental stage. (a) Motor neuron progenitors cultured as 3D spheroids at different stages of development (7, 14 and 21 days) adhered in monolayer culture extend axons over 5 days, e.g. nomenclature ‘7 + 5’ denotes 7 days spheroid culture followed by 5 days adhered monolayer culture. (b) Tuj-1 positive (+) axon length after 5 days adherent culture following seeding at days 7, 14 and 21 days of maturation. Axon length data normalised to day 7 + 5 time-point. (c) Maximum Tuj-1 axon lengths (µm) at each stage of maturation. (d) Percentage (%) axons positive for mature heavy neurofilament SMI-32 after 5 days adherent culture at different development stages. Data indicative of minimum n = 8 per time-point and presented ± standard deviation (SD). **P ≤ 0.01, and ***P ≤ 0.001. Scale bars = 200 µm.