Journal
JOURNAL OF CELL SCIENCE
Volume 133, Issue 12, Pages -Publisher
COMPANY BIOLOGISTS LTD
DOI: 10.1242/jcs.242495
Keywords
Mammary gland; Tissue clearing; Organoid transplantation; Cell dissemination; Atypical PKC iota
Categories
Funding
- Universite de Recherche Paris Sciences et Lettres
- Fondation pour la Recherche Medicale
- Projet Fondation ARC pour la Recherche sur le Cancer [PJA20141201972, PJA20181208073]
- Canceropole Ile de France (EMERG-1, INVADOID 2016)
- Groupement des Entreprises Francaises dans la lutte contre le Cancer, LABoratoires d'EXcellence (LABEX) CellPhyBio [ANR-11LABX-0038, ANR-10-IDEX-0001-02]
- Institut Curie
- Centre National pour la Recherche Scientifique
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Recent developments in techniques for tissue clearing and size reduction have enabled optical imaging of whole organs and the study of rare tumorigenic events in vivo. The adult mammary gland provides a unique model for investigating physiological or pathological processes such as morphogenesis or epithelial cell dissemination. Here, we establish a new pipeline to study rare cellular events occurring in the mammary gland, by combining orthotopic transplantation of mammary organoids with the uDISCO organ size reduction and clearing method. This strategy allows us to analyze the behavior of individually labeled cells in regenerated mammary gland. As a proof of concept, we analyzed the localization of rare epithelial cells overexpressing atypical protein kinase C iota (also known as PRKCI, referred to here as aPKC iota) with an N-terminal eGFP fusion (GFP-aPKC iota(+)) in the normal mammary gland. Using this analytical pipeline, we were able to visualize epithelial aPKC iota(+) cells escaping from the normal mammary epithelium and disseminating into the surrounding stroma. This technical resource should benefit mammary development and tumor progression studies.
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