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Identification of nestin-positive putative mammary stem cells in human breastmilk

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Abstract

Stem cells in mammary tissue have been well characterised by using the mammary stem cell marker, cytokeratin (CK) 5 and the mature epithelial markers CK14, CK18 and CK19. As these markers have never been reported in cells from breastmilk, the aim of this study has been to determine whether mammary stem cells are present in expressed human breastmilk. Cultured cells from human breastmilk were studied by using immunofluorescent labelling and reverse transcription/polymerase chain reaction (RT-PCR). We found a heterogeneous population of cells with differential expression of CK5, CK14, CK18 and CK19. Further, by using the multipotent stem cell marker, nestin, we identified cells in culture that were positive only for nestin or double-positive for CK5/nestin, whereas no co-staining was observed for CK14, CK18 and CK19 with nestin. When cells isolated from breastmilk were analysed by using RT-PCR prior to culture, only nestin and CK18 were detected, thereby indicating that breastmilk contained differentiated epithelial and putative stem cells. Furthermore, fluorescence-activated cell-sorting analysis demonstrated, in breastmilk, a small side-population of cells that excluded Hoechst 33342 (a key property of multipotent stem cells). When stained for nestin, the cells in the side-population were positive, whereas those not in the side-population were negative. The presence of nestin-positive putative mammary stem cells suggests that human breastmilk is a readily available and non-invasive source of putative mammary stem cells that may be useful for research into both mammary gland biology and more general stem cell biology.

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Acknowledgements

The authors are grateful to all the breastfeeding mothers who participated in this research and to the Australian Breastfeeding Association for assistance in their recruitment.

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Authors and Affiliations

  1. School of Biomedical, Biomolecular and Chemical Sciences, Faculty of Life and Physical Sciences, M310, The University of Western Australia, 35 Stirling Highway, Crawley, WA, 6009, Australia

    Mark D. Cregan, Amber Appelbee, Mark L. Brown, John Koppen, Kristin M. E. Piper & Peter E. Hartmann

  2. Department of Obstetrics and Gynaecology, Yong Loo Lin School of Medicine, National University of Singapore, 5 Lower Kent Ridge Road, Singapore, 119074, Singapore

    Yiping Fan, Mahesh A Choolani & Yap-Seng Chong

  3. School of Medicine and Pharmacology, Fremantle Hospital Unit, The University of Western Australia, Alma Street, Fremantle, WA, 6160, Australia

    Borut Klopcic

  4. Medela AG, Lättichstrasse 4b, 6341, Baar, Switzerland

    Leon R. Mitoulas

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  1. Mark D. Cregan
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  2. Yiping Fan
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  3. Amber Appelbee
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  4. Mark L. Brown
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  5. Borut Klopcic
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  6. John Koppen
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  7. Leon R. Mitoulas
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  8. Kristin M. E. Piper
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  9. Mahesh A Choolani
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  10. Yap-Seng Chong
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  11. Peter E. Hartmann
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Corresponding author

Correspondence to Mark D. Cregan.

Additional information

Yiping Fan and Amber Appelbee contributed equally to this work.

This project was funded by Medela, Switzerland.

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Cregan, M.D., Fan, Y., Appelbee, A. et al. Identification of nestin-positive putative mammary stem cells in human breastmilk. Cell Tissue Res 329, 129–136 (2007). https://doi.org/10.1007/s00441-007-0390-x

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  • DOI: https://doi.org/10.1007/s00441-007-0390-x

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