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Preparation and Applications of Organotypic Thymic Slice Cultures
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Immunologia e infezioni
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JoVE Journal Immunologia e infezioni
Preparation and Applications of Organotypic Thymic Slice Cultures

Preparation and Applications of Organotypic Thymic Slice Cultures

DOI:
10.3791/54355-v

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10:10 min

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August 06, 2016

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, ,
1Centre de Recherche,Hôpital Maisonneuve-Rosemont, 2Department of Microbiology, Infectiology and Immunology,Université de Montréal, 3Department of Medicine,Université de Montréal

Capitoli

  • 00:05Titolo
  • 00:45Harvest of Mouse Thymus
  • 01:52Agarose Embedding and Vibratome Sectioning of Thymic Lobes
  • 05:30Preparation of Thymocytes for Overlaying
  • 06:59Overlaying Thymocytes onto the Thymic Slices
  • 07:48Results: Positive and Negative Selection of Thymocytes
  • 09:29Conclusion

Summary

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We describe the preparation of thymic slices that, in combination with flow cytometry, can be used to model positive and negative selection of developing T cells. Thymic slices can also be adapted for the in situ analysis of thymocyte migration, localization, and signaling via immunofluorescence and two-photon microscopy.

Tags

Organotypic Thymic Slice CulturesT Cell DevelopmentPositive And Negative SelectionThymic Cortex And MedullaThymocyte SubsetsThymic MicroenvironmentMouse Thymus DissectionAgarose EmbeddingRPMI-1640 MediaCell Culture Inserts

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