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Generating and Imaging Mouse and Human Epithelial Organoids from Normal and Tumor Mammary Tissue Without Passaging
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1Department of Internal Medicine, Division of Hematology and Oncology,University of Texas Southwestern Medical Center, 2Harold C. Simmons Comprehensive Cancer Center,University of Texas Southwestern Medical Center, 3Department of Molecular Biology,University of Texas Southwestern Medical Center, 4Hamon Center for Regenerative Science and Medicine,University of Texas Southwestern Medical Center, 5Department of Cell Biology,University of Texas Southwestern Medical Center, 6Division of Medical Oncology, Norris Comprehensive Cancer Center, Keck School of Medicine,University of Southern California
Capitoli
- 00:04Introduction
- 00:55Collecting, Digesting Tissue, and Differential Centrifugation
- 04:11Embedding Organoids in BECM and Collagen
- 07:22Results: Immunofluorescent Imaging of Embedded Organoids
- 08:11Conclusion
This protocol discusses an approach for generating epithelial organoids from primary normal and tumor mammary tissue through differential centrifugation. Furthermore, instructions are included for three-dimensional culturing as well as immunofluorescent imaging of embedded organoids.
