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Immunologie et infection
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Isolation of Infiltrating Leukocytes from Mouse Skin Using Enzymatic Digest and Gradient Separation
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Immunologie et infection
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Journal JoVE Immunologie et infection
Isolation of Infiltrating Leukocytes from Mouse Skin Using Enzymatic Digest and Gradient Separation

Isolation of Infiltrating Leukocytes from Mouse Skin Using Enzymatic Digest and Gradient Separation

DOI:
10.3791/53638-v

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07:11 min

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January 25, 2016

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, , ,
1Department of Biology,Macalester College, 2Department of Medicine, Division of Rheumatic and Autoimmune Diseases, Center for Immunology,University of Minnesota

Chapitres

  • 00:05Titre
  • 00:58Flank Skin Harvest and Infiltrating Leukocyte Isolation
  • 04:28Results: Oxazolone (Ox) Challenge Results in Pronounced T Cell and Neutrophil Flank Skin Infiltration
  • 06:17Conclusion

Summary

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Traduction automatique

This protocol describes enzymatic digestion of mouse skin in nutrient-rich medium followed by gradient separation to isolate leukocytes. Cells thus derived can be used for diverse downstream applications. This is an effective, economical, and improved alternative to tissue dissociation machines and harsher trypsin and dispase-based tissue digestion protocols.

Tags

IsolationInfiltrating LeukocytesMouse SkinEnzymatic DigestGradient SeparationCellular MechanismsMolecular MechanismsMouse ModelsSkin PathologiesAtopic DermatitisResident Cell PopulationsSkin HarvestingVisual DemonstrationPrecisionPracticeSurgical ScissorsFlank SkinND4 Swiss-Webster MouseSurgical Blade50 Milliliter Conical TubeHBSSEDTAHEPESFBS Incubation

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