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. 2022 Jan 28;13:562. doi: 10.1038/s41467-021-27895-0

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© The Author(s) 2022

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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Fig. 1 — a Cells from non-lactating tissue (above) and human milk (below) were isolated using either mechanical dissociation and enzymatic digestion or centrifugation, for downstream analysis. b Mammary cells from both non-lactating breast tissue (top) or lactating milk cells (bottom) were cultured either in i 2D (n = 10, see Supplementary Fig. 2) or ii 3D, scale bar represents 250 μm. c Representative flow cytometric profiles of immune/stromal (Draq5⁺/CD45⁺), luminal (Draq5⁺/EpCAM⁺/CD49f^+/−) and myoepithelial cells (Draq5⁺/EpCAM⁻/CD49f⁺) from i NMCs and ii LMCs. d Schematic diagram for the scRNA-seq experimental set-up for cell samples from seven non-lactating participants and nine lactating females. e Uniform manifold approximation and projection (UMAP) dimensional reduction of the mammary cells reveals distinct clusters arising from NMCs and LMCs.