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Isolation of Lamina Propria Mononuclear Cells from Murine Colon Using Collagenase E
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1Department of Pediatrics, Division of Hematology / Oncology and Bone Marrow Transplantation,University of Miami Miller School of Medicine, 2Batchelor Children’s Research Institute,University of Miami Miller School of Medicine, 3Department of Microbiology & Immunology,University of Miami Miller School of Medicine, 4Sylvester Comprehensive Cancer Center,University of Miami Miller School of Medicine, 5Holtz Children’s Hospital,University of Miami Miller School of Medicine
Chapters
- 00:04Title
- 01:00Harvesting the Colon
- 01:32Cleaning the Colon
- 03:10Collagenase Digestion 1
- 03:34Silica-Based Density Separation Media Gradient Separation
- 05:44Collect Mononuclear Cells from the Gradient Interface
- 06:39Results: Isolation of Lamina Propria Mononuclear Cells
- 08:40Conclusion
The goal of this protocol is to isolate mononuclear cells that reside in the lamina propria of the colon by enzymatic digestion of the tissue using collagenase. This protocol allows for the efficient isolation of mononuclear cells resulting in a single cell suspension which in turn can be used for robust immunophenotyping.
