Organoid cultures from normal and cancer-prone human breast tissues preserve complex epithelial lineages

Jennifer M Rosenbluth,Carman Man-Chung Li,Andrea Richardson,Ron C J Schackmann,Norman Sachs,Hendrik J Kuiken,Mackenzie Boedicker,Jane Brock,Judy Garber,Joan S Brugge,Deborah Dillon,G Kenneth Gray,Hans Clevers,Laura M Selfors

doi:10.1038/s41467-020-15548-7

Abstract

Recently, organoid technology has been used to generate a large repository of breast cancer organoids. Here we present an extensive evaluation of the ability of organoid culture technology to preserve complex stem/progenitor and differentiated cell types via long-term propagation of normal human mammary tissues. Basal/stem and luminal progenitor cells can differentiate in culture to generate mature basal and luminal cell types, including ER+ cells that have been challenging to maintain in culture. Cells associated with increased cancer risk can also be propagated. Single-cell analyses of matched organoid cultures and native tissues by mass cytometry for 38 markers provide a higher resolution representation of the multiple mammary epithelial cell types in the organoids, and demonstrate that protein expression patterns of the tissue of origin can be preserved in culture. These studies indicate that organoid cultures provide a valuable platform for studies of mammary differentiation, transformation, and breast cancer risk.

Highlights

Organoid technology has been used to generate a large repository of breast cancer organoids
Sachs et al developed organoid culture conditions optimized for long term in vitro propagation of breast tumor and normal epithelium[4]
This study demonstrates the ability of CyTOF technology to provide a high resolution analysis of the extent to which organoid cultures faithfully preserve subtypes of epithelial cells from the normal mammary gland

Summary

Introduction

Organoid technology has been used to generate a large repository of breast cancer organoids. Single-cell analyses of matched organoid cultures and native tissues by mass cytometry for 38 markers provide a higher resolution representation of the multiple mammary epithelial cell types in the organoids, and demonstrate that protein expression patterns of the tissue of origin can be preserved in culture These studies indicate that organoid cultures provide a valuable platform for studies of mammary differentiation, transformation, and breast cancer risk. Recent studies of murine models as well as human tissues have suggested far greater complexity of mammary epithelial hierarchy, and have identified specific differences in the composition of these epithelial compartments associated with pregnancy, with aging, and with inherited mutations in BC predisposition genes[10,11,12,13,14] These studies and others suggest a high level of complexity within the mammary epithelium, with one or more cell types contributing to the development of BC. Organoid technology allows the growth and characterization of multiple normal mammary epithelial cell lineages in a single culture, which will enable a greater understanding of the genesis of different BC subtypes

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Results

Conclusion