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. 2019 Jun 20;9:8921. doi: 10.1038/s41598-019-45407-5

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© The Author(s) 2019

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fabrication of spheroids consisting of human neurons (hNs) and/or human Schwann cells (hSCs) after 2 days in vitro. Presence of hSCs expedited spheroid formation in the co-culture system while the hN-only condition did not form spheroids in 2 days. hSC spheroids were fabricated with three different cellular densities: 25,000 (a) 50,000 (b) and 75,000 (c). Co-culture spheroids were created with a constant hN density (75,000) but a changing hSC density: 25,000 (d) 50,000 (e) and 75,000. (f) In parallel, hN spheroids were fabricated at three different densities: 50,000 (g), 75,000 (h) and 100,000. (i,j) Comparison of diameters of different spheroids revealed that co-culture spheroids were more compact as compared to mono-culture spheroids, showing that the affinity of hSCs for hNs resulted in a more tightly packed cluster of cells. K (multiple of thousand). N = 4 and error bars represent standard error of the mean (SEM). Scale bar: 100 µm. (****p-value ≤ 0.0001), (***p-value ≤ 0.001), (**p-value ≤ 0.01), (*p-value ≤ 0.05).