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Title: An optimized method for establishing high purity murine CD8+ T cell cultures.
Austin Authors: Zanker, Damien;Xiao, Kun;Oveissi, Sara;Guillaume, Philippe;Luescher, Immanuel F;Chen, Weisan
Affiliation: T Cell Laboratory, Ludwig Institute for Cancer Research, Austin Health, Heidelberg, Victoria, Australia 3084, Australia
Issue Date: 22-Oct-2012
Publication information: Journal of Immunological Methods 2012; 387(1-2): 173-80
Abstract: 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.
Gov't Doc #: 23098837
DOI: 10.1016/j.jim.2012.10.012
Type: Journal Article
Subjects: Amino Acid Sequence
Antigens, CD45.immunology.metabolism
Apoptosis.drug effects.immunology
CD8-Positive T-Lymphocytes.drug effects.immunology.metabolism
Cell Culture Techniques.methods
Cell Proliferation.drug effects
Cells, Cultured
Dose-Response Relationship, Drug
Lymphocyte Count
Mice, Inbred BALB C
Mice, Inbred C57BL
Appears in Collections:Journal articles

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