title: Cytokines and culture medium have a major impact on human in vitro T-cell differentiation.
abstract: An important proof of principle has been achieved with the development of an in vitro T-cell differentiation assay based on the coculture of hematopoietic progenitors with the OP9-Delta1 stromal cell line. The original murine T cell differentiation assay has since been adapted for human T-cell differentiation, however with lower efficiency. The choice of both medium and cytokines is crucial in this assay, therefore our work has been focused on these two factors. The use of freshly reconstituted medium, the optimization of interleukine-7 (IL-7) concentration, and the addition of stem cell factor (SCF) have allowed to improve the proliferation of progenitors and T-cell precursors as well as the yield of double positive CD4+CD8+ T cells, and mature γδ and αβ T cells. These optimizations make the OP9-Delta1 system sensitive enough to perform both quantitative and qualitative assays with various type of progenitors, including those transduced by a retroviral vector. The improved OP9-Delta1 assay therefore constitutes an extremely useful test for basic research purposes and for translational medicine.
authors.affiliation: INSERM U768, Université Paris-Descartes, Faculté de Médecine René Descartes, Paris, France.
authors.forename: Emmanuelle M
authors.initials: EM
authors.lastname: Six
authors.forename: Fatine
authors.initials: F
authors.lastname: Benjelloun
authors.forename: Alexandrine
authors.initials: A
authors.lastname: Garrigue
authors.forename: Delphine
authors.initials: D
authors.lastname: Bonhomme
authors.forename: Estelle
authors.initials: E
authors.lastname: Morillon
authors.forename: Julien
authors.initials: J
authors.lastname: Rouiller
authors.forename: Laure
authors.initials: L
authors.lastname: Cacavelli
authors.forename: Johanna
authors.initials: J
authors.lastname: Blondeau
authors.forename: Kheira
authors.initials: K
authors.lastname: Beldjord
authors.forename: Salima
authors.initials: S
authors.lastname: Hacein-Bey-Abina
authors.forename: Marina
authors.initials: M
authors.lastname: Cavazzana-Calvo
authors.forename: Isabelle
authors.initials: I
authors.lastname: André-Schmutz
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D016207:Cytokines / Q000276:immunology / Q000494:pharmacology*; D006412:Hematopoietic Stem Cells / Q000276:immunology; D006801:Humans; D015851:Interleukin-7 / Q000494:pharmacology; D019089:Stem Cell Factor / Q000494:pharmacology; D017154:Stromal Cells / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology* / Q000187:drug effects*
chemical_list: D000276:Adjuvants, Immunologic; D017077:Culture Media, Conditioned; D016207:Cytokines; D015851:Interleukin-7; D019089:Stem Cell Factor