Protocol to generate mouse oviduct epithelial organoids for viral transduction and whole-mount 3D imaging

Matthew J Ford; Keerthana Harwalkar; Yojiro Yamanaka

doi:10.1016/j.xpro.2022.101164

Protocol to generate mouse oviduct epithelial organoids for viral transduction and whole-mount 3D imaging

STAR Protoc. 2022 Feb 9;3(1):101164. doi: 10.1016/j.xpro.2022.101164. eCollection 2022 Mar 18.

Authors

Matthew J Ford¹, Keerthana Harwalkar¹, Yojiro Yamanaka¹

Affiliation

¹ Rosalind and Morris Goodman Cancer Institute, McGill University, Department of Human Genetics, 1160 Pine Avenue West, Montreal, QC H3A 1A3, Canada.

Abstract

Epithelial cells lining the oviduct/fallopian tube are essential in reproduction and have been identified as the cell-of-origin in high-grade serous ovarian carcinoma (HGSOC). This protocol describes the generation of organoids from mouse oviduct epithelial cells, providing a powerful in vitro tool to study epithelial homeostasis and malignant transformation. We also outline a protocol for whole-mount immunofluorescence and 3D confocal imaging. In addition, we describe approaches of viral transduction to investigate gene function in organoid development and epithelial cell behavior. For complete details on the use and execution of this profile, please refer to Ford et al. (2021).

Keywords: Cancer; Cell Biology; Cell culture; Cell isolation; Developmental biology; Microscopy; Organoids.

Publication types

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

MeSH terms

Animals
Epithelial Cells / pathology
Fallopian Tubes / diagnostic imaging
Female
Humans
Imaging, Three-Dimensional*
Mice
Organoids* / physiology
Oviducts