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. 2023 Nov 22;14:7639. doi: 10.1038/s41467-023-43239-6

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© The Author(s) 2023

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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Fig. 1 — a Schematic of the formation of MSC aggregates via acoustofluidic assembly and N-cadherin mediated cell–cell interactions to enhance the MSC secretome. b The simulation results showed that a three-dimensional circular vortex tube generated by a high-intensity focal point at the center of the substrate functioned as a virtual wall, trapping and aggregating cells at the center of the droplet. c Illustration of the comparison between 2D monolayer MSC culture and acoustofluidic 3D MSC aggregation. d Fluorescence images of MSCs before/after the acoustofluidic assembly, and the morphology of MSC aggregates after 3 days of incubation. The green color indicates alive cells. The blue color indicates cell nuclei, and the red color indicates the cytoskeleton. n = 4 tests with similar results. Scale bar: 100 μm. e CyQUANT™ Cell Proliferation Assay measuring the DNA contents of different groups. The statistical analysis was performed using one-way ANOVA with Tukey’s post-hoc test. Data are graphed as the mean ± SD (n = 4, biological repeats). Source data are provided as a Source Data file.