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Abstract

Nanoscale Imaging of Human Milk Cells †

by
Qiongxiang Lin
1,2,
Sharon L. Perrella
1,3,4,
Ashleigh H. Warden
1,3,4,
Cameron W. Evans
1,2,
Donna T. Geddes
1,2,3,4,
Leon R. Mitoulas
1,2,5,
Haibo Jiang
1,2,6,
Kai Chen
2,7 and
Killugudi Swaminatha Iyer
1,2,*
1
School of Molecular Sciences, The University of Western Australia, Crawley, WA 6009, Australia
2
ARC Training Centre for Next-Gen in Biomedical Analysis, School of Molecular Sciences, The University of Western Australia, Perth, WA 6009, Australia
3
ABREAST Network, Perth, WA 6000, Australia
4
UWA Centre for Human Lactation Research and Translation, Crawley, WA 6009, Australia
5
Medela AG, ZUG 6340 Baar, Switzerland
6
Department of Chemistry, The University of Hong Kong, Pok Fu Lam, Hong Kong 999077, China
7
School of Biomedical Sciences, The University of Western Australia, Crawley, WA 6009, Australia
*
Author to whom correspondence should be addressed.
†
Presented at Australian Breastfeeding + Lactation Research and Science Translation Conference (ABREAST Conference 2024), Perth, Australia, 15 November 2024.
Proceedings 2025, 112(1), 23; https://doi.org/10.3390/proceedings2025112023
Published: 27 February 2025
(This article belongs to the Proceedings of Australian Breastfeeding + Lactation Research and Science Translation Conference 2024)
Versions Notes

Abstract

:
Human milk is a complex biofluid containing a diverse array of cells crucial for infant health. Despite their importance, our understanding of these cells remains incomplete due to technical challenges. To fully comprehend human milk cells, high-resolution imaging technologies that can directly measure biological processes are required. We have developed a specialized imaging platform combining light and electron microscopy for human milk cell imaging. To identify different cell types, human milk cells were first stained with several specific cell markers (e.g., EpCAM and MUC1 for lactocytes, CD16 and CD66b for neutrophils, and HLA-DR and CD68 for macrophages) prior to light (confocal) microscopy. Following this, the same cells were processed with osmium staining, resin embedding, and sectioning for electron microscopy, allowing us to observe ultrastructural details. Our imaging workflow has enabled nanoscale visualization of human milk cells, resulting in a first-of-its-kind comprehensive database profiling the organelle-level ultrastructure of different cell types present in human milk. The cells in the human milk are highly heterogenous, featuring a large proportion of lactocytes and lipid droplets, binucleated lactocytes, neutrophil aggregation, neutrophil extracellular traps, dendritic cells/macrophages with bacteria, and immunophagocytosis. This study provides valuable cellular insights contributing to a deeper understanding of human milk biology.

Author Contributions

Conceptualization, K.S.I., K.C. and H.J.; methodology, Q.L., K.S.I., K.C., H.J., C.W.E., D.T.G., S.L.P. and A.H.W.; formal analysis, Q.L. and K.C.; investigation, A.H.W. and S.L.P.; resources, K.S.I., D.T.G. and L.R.M.; writing—original draft preparation, Q.L.; writing—review and editing, K.S.I., K.C., H.J., C.W.E., D.T.G., L.R.M., S.L.P. and A.H.W.; supervision, C.W.E. and K.S.I.; funding acquisition, K.S.I. All authors have read and agreed to the published version of the manuscript.

Funding

This research was funded by the Australian Research Council (ARC) (Project ID: IC210100056). Q.L. is supported by the Research Training Program Fees Offset Scholarship and ARC On-Water Electrochemistry HDR Scholarship. D.T.G., S.L.P., and A.H.W. are supported by an unrestricted research grant from Medela AG (Switzerland), administered by The University of Western Australia. L.R.M. is supported by Medela AG. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

Institutional Review Board Statement

The study was conducted in accordance with the Declaration of Helsinki and approved by the UWA Human Research Ethics Committee (2019/RA/4/20/6498; the date of approval 21/02/2024).

Informed Consent Statement

Informed consent was obtained from all participants involved in the study.

Acknowledgments

We thank all the participants who donated breast milk for the study.

Conflicts of Interest

D.T.G. declares past participation in the Scientific Advisory Board of Medela. AG. D.T.G., S.L.P. and A.H.W. are supported by an unrestricted research grant from Medela AG, administered by The University of Western Australia. L.R.M. is supported by Medela AG. The funder had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results. All other authors declare no conflicts of interest.
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content.

Share and Cite

MDPI and ACS Style

Lin, Q.; Perrella, S.L.; Warden, A.H.; Evans, C.W.; Geddes, D.T.; Mitoulas, L.R.; Jiang, H.; Chen, K.; Iyer, K.S. Nanoscale Imaging of Human Milk Cells. Proceedings 2025, 112, 23. https://doi.org/10.3390/proceedings2025112023

AMA Style

Lin Q, Perrella SL, Warden AH, Evans CW, Geddes DT, Mitoulas LR, Jiang H, Chen K, Iyer KS. Nanoscale Imaging of Human Milk Cells. Proceedings. 2025; 112(1):23. https://doi.org/10.3390/proceedings2025112023

Chicago/Turabian Style

Lin, Qiongxiang, Sharon L. Perrella, Ashleigh H. Warden, Cameron W. Evans, Donna T. Geddes, Leon R. Mitoulas, Haibo Jiang, Kai Chen, and Killugudi Swaminatha Iyer. 2025. "Nanoscale Imaging of Human Milk Cells" Proceedings 112, no. 1: 23. https://doi.org/10.3390/proceedings2025112023

APA Style

Lin, Q., Perrella, S. L., Warden, A. H., Evans, C. W., Geddes, D. T., Mitoulas, L. R., Jiang, H., Chen, K., & Iyer, K. S. (2025). Nanoscale Imaging of Human Milk Cells. Proceedings, 112(1), 23. https://doi.org/10.3390/proceedings2025112023

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