Ex Vivo Organoid Model of Adenovirus-Cre Mediated Gene Deletions in Mouse Urothelial Cells

Overview

This protocol describes a rapid and efficient method for generating bladder cancer organoids from mouse urothelial cells through ex vivo gene editing. The approach allows for quick characterization of organoids with specific gene deletions.

Key Study Components

Area of Science

• Neuroscience
• Cell Biology
• Oncology

Background

• Bladder cancer organoids can be generated from normal mouse urothelial cells.
• Traditional methods involve extensive mouse breeding, which is time-consuming.
• Ex vivo gene editing provides a faster alternative.
• Adenovirus transduction is utilized for efficient gene deletion.

Purpose of Study

• To develop a protocol for generating bladder cancer organoids.
• To facilitate the study of gene function in bladder cancer.
• To reduce the time required for organoid generation.

Methods Used

• Harvesting mouse urothelial cells.
• Ex vivo transduction with adenovirus driving Cre expression.
• Characterization of organoids in vitro and in vivo.
• Use of sterile instruments and reagents for dissection and culture.

Main Results

• Successful generation of bladder cancer organoids from edited cells.
• Efficient adenovirus transduction leading to gene deletions.
• Characterization of organoids confirmed their viability and relevance.
• Reduction in workflow time compared to traditional breeding methods.

Conclusions

• The protocol provides a quick and efficient method for organoid generation.
• Ex vivo gene editing is a valuable tool for cancer research.
• This approach can accelerate the study of bladder cancer biology.

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