An optimized method for establishing high purity murine CD8+ T cell cultures.

Détails

ID Serval
serval:BIB_0F323C9E03B5
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
An optimized method for establishing high purity murine CD8+ T cell cultures.
Périodique
Journal of Immunological Methods
Auteur(s)
Zanker D., Xiao K., Oveissi S., Guillaume P., Luescher I.F., Chen W.
ISSN
1872-7905 (Electronic)
ISSN-L
0022-1759
Statut éditorial
Publié
Date de publication
2013
Volume
387
Numéro
1-2
Pages
173-180
Langue
anglais
Résumé
Establishing CD8(+) T cell cultures has been empirical and the published methods have been largely individual laboratory based. In this study, we optimized culturing conditions and show that IL-2 concentration is the most critical factor for the success of establishing CD8(+) T cell cultures. High IL-2 concentration encouraged T cells to non-specifically proliferate, express a B cell marker, B220, and undergo apoptosis. These cells also lose typical irregular T cell morphology and are incapable of sustaining long-term cultures. Using tetramer and intracellular cytokine assessments, we further demonstrated that many antigen-specific T cells have been rendered nonfunctional when expanded under high IL-2 concentration. When IL-2 is used in the correct range, B220-mediated cell depletion greatly enhanced the success rate of such T cell cultures.
Mots-clé
Amino Acid Sequence, Animals, Antigens, CD45/immunology, Antigens, CD45/metabolism, Apoptosis/drug effects, Apoptosis/immunology, CD8-Positive T-Lymphocytes/drug effects, CD8-Positive T-Lymphocytes/immunology, Cell Culture Techniques/methods, Cell Proliferation/drug effects, Cells, Cultured, Dose-Response Relationship, Drug, Interferon-gamma/immunology, Interferon-gamma/metabolism, Interleukin-2/pharmacology, Lymphocyte Count, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL
Pubmed
Web of science
Création de la notice
12/04/2013 18:00
Dernière modification de la notice
20/08/2019 12:36
Données d'usage