Generation and functional characterization of murine mammary organoids

Hon Yan Kelvin Yip; Antonella Papa

doi:10.1016/j.xpro.2021.100765

Generation and functional characterization of murine mammary organoids

STAR Protoc. 2021 Aug 24;2(3):100765. doi: 10.1016/j.xpro.2021.100765. eCollection 2021 Sep 17.

Authors

Hon Yan Kelvin Yip¹, Antonella Papa¹

Affiliation

¹ Cancer Program, Monash Biomedicine Discovery Institute and Department of Biochemistry and Molecular Biology, Monash University, Melbourne, VIC 3800, Australia.

Abstract

3D cultures of mammary epithelial cells purified from murine models provide a unique resource to study genetically defined breast cancer and response to targeted therapies. Here, we describe step-by-step experimental procedures for the successful establishment of murine mammary organoid lines isolated from mammary glands or mammary tumors driven by mutations in components of the PI3K pathway. These detailed protocols also include procedures to perform assays that can be adopted to screen response to drug treatments and to inform better therapies. For details on potential applications and use of this protocol, please refer to Yip et al. (2020).

Keywords: Cancer; Cell culture; Cell isolation; Cell-based Assays; Organoids.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Death / physiology
Cryopreservation
Female
Mammary Glands, Animal / cytology*
Mammary Glands, Animal / physiology
Mammary Neoplasms, Experimental / genetics
Mammary Neoplasms, Experimental / pathology*
Metabolic Networks and Pathways
Mice
Mice, Inbred C57BL
Organ Culture Techniques / instrumentation
Organ Culture Techniques / methods*
Organoids*
Phosphatidylinositol 3-Kinases / genetics*
Phosphatidylinositol 3-Kinases / metabolism