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Isolamento de células mononucleares de lamina propria de cólon murino usando 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
O objetivo deste protocolo é isolar as células mononucleares que residem na lâmina própria do cólon por digestão enzimática do tecido usando Collagenase. Este protocolo permite a isolação eficiente de pilhas mononucleares tendo por resultado uma única suspensão da pilha que por sua vez possa ser usada para immunophenotyping robusto.
