Characterization of organoid cultured human breast cancer

Nadine Goldhammer,Ole William Petersen,Vera Timmermans-Wielenga,Jiyoung Kim

doi:10.1186/s13058-019-1233-x

Nadine Goldhammer, Ole William Petersen + Show 2 more

Open Access

https://doi.org/10.1186/s13058-019-1233-x

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Abstract

Organoid cultures are increasingly used to model human cancers experimentally with a view to tailoring personalized medicine and predicting drug responses. Breast cancer is no exception, but in particular, primary breast cancer poses some inherent difficulties due to the frequent presence of residual non-malignant cells in the biopsies. We originally developed an assay for the distinction between malignant and non-malignant structures in primary breast cancer organoid cultures (Petersen et al., Proc Natl Acad Sci (USA) 89(19):9064–8, 1992). Here, we apply this assay to assess the frequency of normal-like organoids in primary breast carcinoma cultures and the cellular composition as a consequence of passaging. We find that in consecutively collected samples of primary human breast cancers, residual non-malignant tissues were observed histologically in five out of ten biopsies. Based on relevant morphogenesis and correct polarization as recorded by expression in luminal epithelial cells of mucin 1 (Muc1), occludin, and keratin 19 (K19) and expression in basal cells of integrin β4, p63, and K14, non-malignant organoids were present in all primary human breast cancer-derived cultures. Furthermore, passaging in a contemporary culture medium was in favor of the selective expansion of basal-like cells. We conclude that organoid cultures of human breast cancers are most representative of the tissue origin in primary culture.

Highlights

Organoid culture conditions have been devised for multiple human cancer types including those of the colon, esophagus, pancreas, stomach, liver, endometrium, prostate, and breast
We initially aimed for optimal separation of normal luminal epithelial and myoepithelial cells by a FACSbased protocol relying on a combination of antibodies against the EpCAM family-related trophoblast surface antigen 2, Trop2, and the nerve growth factor receptor, CD271 (Fig. 1)
Upon plating inside the breast cancer organoid assay as single cells, luminal cells grew up to form approximately 50-μm-sized acinuslike structures before growth arrest while myoepithelial cells formed larger ball-like structures, both reminiscent of what has been described originally with another culture medium (Fig. 1 and [8])

Summary

Introduction

Organoid culture conditions have been devised for multiple human cancer types including those of the colon, esophagus, pancreas, stomach, liver, endometrium, prostate, and breast (for review, see [1]). A recipe based on a key addition of Neuregulin-1 to the culture medium allowed for the culture of > 80% of human breast cancer biopsies. In a preliminary characterization of the breast cancer organoids, some were passaged for > 20 passages [2]. A total of 95 “lines” were obtained, but in general, the identity of the cultured cells was not obvious. For example, predict drug response by using 2D or 3D primary cell culture models of cancer have been complicated by the potential concurrent growth of cells from residual, non-malignant

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