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Naïve T Lymphocyte Infection by Murine Stem Cell Virus (MSCV) Protocol

Jia Li Jia Li

Published: Nov 20, 2011 DOI: 10.21769/BioProtoc.156 Views: 12704

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Abstract

This assay can be used for studying genes related to lymphocyte proliferation and differentiation in vitro. Stimulating lymphocytes to proliferate is important for the infection efficiency of MSCV. After overnight culture, if naïve cells proliferate extensively, the following spin infection will have high efficiency. After infection, cells are ready for the following assay as they will be undergoing proliferation or differentiation.

Background

Materials and Reagents

  1. Mouse
  2. Fetal bovine serum (FBS)
  3. Anti-CD3 and Anti-CD28 antibody (Ab) (Biolegend, catalog number: 100302 , 102102 ）
  4. IL-2 (R&D systems, catalog number: 402-ML-020 )
  5. Polybrene (Hexadimethrine bromide) (Sigma-Aldrich, catalog number: H9268 )
  6. Fetal bovine serum (FBS) (Gemini Bio-Products, catalog number: 900-108 )
  7. Penicillin/streptomycin solution (Life Technologies, Gibco®, catalog number: 15140-122 )
  8. 2-mercaptoethanol (Life Technologies, InvitrogenTM, catalog number: 21985-023 )
  9. PBS solution (1x phosphate buffered saline) (Life Technologies, Gibco®, catalog number: 10010-023 )
  10. MSCV (Clontech, catalog number: 634401 )
  11. RPMI-1640 medium (Life Technologies, Gibco®, catalog number: 11875-093 ) (see Recipes)

Equipment

  1. Centrifuges
  2. 24-well plate
  3. Water bath
  4. Parafilm

Procedure

Copyright: © 2011 The Authors; exclusive licensee Bio-protocol LLC.

Category

Immunology > Immune cell function > Lymphocyte

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