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. 2022 Dec 21;13:529. doi: 10.1186/s13287-022-03215-1

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© The Author(s) 2022

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 3 — Chambered cardiac organoids are composed of cell types from the various cardiac lineages. A Uniform manifold approximation and projection (UMAP) of all cells, representing distinct identities of cells in day 8, day 14 and day 21 CCOs (top). UMAP revealed ten main cell clusters within the three time points (bottom). B Gene expression levels of the identified seven clusters related to the cardiac lineage are isolated in this heatmap. Each cluster identified is expressing their own marker genes. The singular endothelial cell cluster contains cells from all three timepoints of the organoids. C Representative confocal immunofluorescence image showing expression of cardiomyocyte, smooth muscle cell and endothelial cell lineage specific markers in the dissociated CCOs. Scale bars, 100 µm. D–E Quantitative RT-qPCR gene expression comparison of 2D cultured cardiomyocytes and 3D cardiac organoids of day 14 (D) and day 21 (E), normalized to day 7 cardiomyocytes. Genes shown represent markers of cardiomyocyte, smooth muscle and endothelial cell lineages (n = 3; mean ± s.e.m; housekeeping gene, ACTB; ***P < 0.001; **P < 0.01; *P < 0.05; two-tailed Student’s t test)