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. 2021 Jan 22;11:2080. doi: 10.1038/s41598-021-81335-z

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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/.

PMC Copyright notice

Motor neuron progenitors form 3D networks in the OrganoPlate. (A) Image of an OrganoPlate 3-lane. The OrganoPlate is a microfluidic platform based on a 384-well plate design containing 40 individual tissue culture chips. The image is copyright of MIMETAS BV. (B) Each tissue culture chip contains 3 microfluidic lanes that join in the center of the chip. The 3-lane architecture in the chips’ center is assured by phaseguides that enable patterning of extracellular matrix gel. Inlet and outlet wells allow access to each lane: top inlet (TI) and top outlet (TO) wells allow access to top lane; middle inlet (MI) and outlet (MO) wells to middle lane; and bottom inlet (BI) and outlet (BO) wells to the bottom lane. Matrigel-GFR without cells is added to the middle lane. Following gelation, motor neuron progenitors embedded in matrigel-GFR are seeded in the top lane. (C) Schematic overview of cell seeding procedure. Matrigel-GFR is seeded in the middle lane and MNPs embedded in matrigel-GFR are seeded in the top lane (panel 1). MNPs form networks in the top lane (panel 2), after which neurites protrude into the middle and eventually bottom lane (panel 3). Blue color indicates presence of gel, red color indicates presence of medium. (D) Schematic overview of the spatial segregation of motor neuron somata (top lane) and axons (middle and bottom lanes) in OrganoPlate 3-lane cultures.