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Immunologie et infection
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Generation of Induced Regulatory T Cells from Primary Human Naïve and Memory T Cells
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Immunologie et infection
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Journal JoVE Immunologie et infection
Generation of Induced Regulatory T Cells from Primary Human Naïve and Memory T Cells

Generation of Induced Regulatory T Cells from Primary Human Naïve and Memory T Cells

DOI:
10.3791/3738-v

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14:23 min

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April 16, 2012

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, , , ,
1Department of Microbiology, Immunology and Molecular Genetics,University of Kentucky

Chapitres

  • 00:05Titre
  • 02:22Isolation of Human Peripheral Blood Mononuclear Cells (PBMCs) from Buffy Coat
  • 03:58Magnetic Negative Selection of T Cells from PBMCs using EasySep Enrichment Kit
  • 06:16Cell Culture Conditions to Induce Regulatory T Cells
  • 07:11Fluorescence-activated Cell Sorting of Three Populations of T Cells
  • 08:28Suppression Assay
  • 10:32Representative Sorting and Suppression Assay Results
  • 13:14Conclusion

Summary

Traduction automatique

Traduction automatique

We describe a method for generating regulatory, memory and naïve T cells from a single human blood donor. Polarized Tregs can be then compared to other subsets in a variety of genetic and functional applications with genetic homogeneity, including a suppression assay also detailed here.

Tags

Induced Regulatory T CellsPrimary Human Naive T CellsMemory T CellsImmunosuppressive CD4+ Regulatory T CellsPeripheral ToleranceImmunologic HomeostasisSelf AntigensCommensal AntigensTreg BiologyTherapeutic Injection Of TregsMurine ModelsColitisType I DiabetesRheumatoid ArthritisGraft Versus Host DiseaseEx Vivo ExpansionPeripheral Blood Mononuclear CellsCD4+ T CellsIn Vitro Cell CultureFluorescence Activated Cell Sorting (FACS)Forkhead Box P3 (FoxP3)Activation-induced Transcription Factor

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