3D in vitro culture system to study collective migration in mammary organoid epithelium

Yunzhe Lu; Ruolan Deng; Huanyang You; Pengfei Lu

STAR Protocols (Sep 2021)

3D in vitro culture system to study collective migration in mammary organoid epithelium

Yunzhe Lu,
Ruolan Deng,
Huanyang You,
Pengfei Lu

Affiliations

Yunzhe Lu: School of Life Science and Technology, ShanghaiTech University, Shanghai 201210, China; Corresponding author
Ruolan Deng: School of Life Science and Technology, ShanghaiTech University, Shanghai 201210, China; University of Chinese Academy of Sciences, Beijing, China; Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai, China
Huanyang You: School of Life Science and Technology, ShanghaiTech University, Shanghai 201210, China; University of Chinese Academy of Sciences, Beijing, China; Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences, Shanghai, China
Pengfei Lu: School of Life Science and Technology, ShanghaiTech University, Shanghai 201210, China; Corresponding author

Journal volume & issue: Vol. 2, no. 3
p. 100778

Abstract

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Summary: We recently established an in vitro culture system in which mammary gland organoid undergoes directional migration in response to an FGF10 concentration gradient. Here, we describe a step-by-step protocol for preparing organoids, the setup of the 3D culture system, and the image acquisition approach. The technical difficulties in conducting the 3D migration assay are choosing epithelial organoids of appropriate sizes and manually paring organoids and beads pre-soaked in FGF10 within a desirable distance (∼100 μm).For complete details on the use and execution of this protocol, please refer to Lu et al. (2020).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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