A reproducible scaffold-free 3D organoid model to study neoplastic progression in breast cancer

Journal of Cell Communication and Signaling

article

Djomehri, Sabra I.¹  Burman, Boris¹  Gonzalez, Maria E.¹  Takayama, Shuichi⁴  Kleer, Celina G.¹ 

[1] Department of Pathology, University of Michigan Medical School;Molecular and Cellular Pathology Training Program, University of Michigan;Rogel Cancer Center, University of Michigan;Department of Biomedical Engineering, Biointerfaces Institute, University of Michigan;Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology

关键词: 3D cell culture;    Organoids;    Hanging drop;    Human breast cancer;    Morphogenesis;    Phenotype;    Microenvironment;    EMT;    Mesenchymal stromal cells;    Metaplastic;    CCN6;   

DOI  :  10.1007/s12079-018-0498-7

学科分类：分子生物学，细胞生物学和基因

来源: Springer

【 摘 要 】

While 3D cellular models are useful to study biological processes, gel-embedded organoids have large variability. This paper describes high-yield production of large (~1 mm diameter), scaffold-free, highly-spherical organoids in a one drop-one organoid format using MCF10A cells, a non-tumorigenic breast cell line. These organoids display a hollow lumen and secondary acini, and express mammary gland-specific and progenitor markers, resembling normal human breast acini. When subjected to treatment with TGF-β, the hypoxia-mimetic reagent CoCl2, or co-culture with mesenchymal stem/stromal cells (MSC), the organoids increase collagen I production and undergo large phenotypic and morphological changes of neoplastic progression, which were reproducible and quantifiable. Advantages of this scaffold-free, 3D breast organoid model include high consistency and reproducibility, ability to measure cellular collagen I production without noise from exogenous collagen, and capacity to subject the organoid to various stimuli from the microenvironment and exogenous treatments with precise timing without concern of matrix binding. Using this system, we generated organoids from primary metaplastic mammary carcinomas of MMTV-Cre;Ccn6fl/fl mice, which retained the high grade spindle cell morphology of the primary tumors. The platform is envisioned to be useful as a standardized 3D cellular model to study how microenvironmental factors influence breast tumorigenesis, and to potential therapeutics.

【 授权许可】

CC BY