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Optimization of Human Th17 Cell Differentiation in Vitro: Evaluating Different Polarizing Factors Publisher Pubmed



Ganjalikhani Hakemi M1, 2 ; Ghaedi K3 ; Andalib A1 ; Hosseini M4 ; Rezaei A1, 2
Authors
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Authors Affiliations
  1. 1. Immunology Dept., Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
  2. 2. Applied Physiology Research Center, Faculty of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
  3. 3. Biology Dept., Faculty of Sciences, University of Isfahan, Isfahan, Iran
  4. 4. Epidemiology Dept., Faculty of Health, Isfahan University of Medical Sciences, Isfahan, Iran

Source: In Vitro Cellular and Developmental Biology - Animal Published:2011


Abstract

Regarding discrepancies that exist among different studies which have tried to clarify critical factors in human Th17 cell differentiation, the aim of this study was to identify the best condition for human Th17 differentiation and to clarify the possible role of TGF-β in differentiation of these cells. Naive CD4 + T cells were isolated from cord blood samples and cultured either in X-VIVO 15 serum-free medium or RPMI 1640 containing 10% FBS. Purified cells were treated with different combinations of polarizing cytokines (TGF-β, IL-1β, IL-6, IL-23 and IL-21) followed by analysis of the expression of characteristic genes and their relevant cytokines by real-time quantitative RT-PCR and ELISA method, respectively. Our data indicate that a combination of TGF-β plus IL-6 and IL-23 cytokines in X-VIVO 15 serum-free medium could be applied as the best condition for developing human Th17 cells in compare with other studied cytokine treatments. It is shown that TGF-β could be considered as a positive regulator for human Th17 cell differentiation only if applied in average concentrations. Interestingly, polarizing treatments in absence of TGF-β, induced double-secreting Th17 cells which co-express IL-17 and IFN-γ whereas polarization in presence of TGF-β-induced single-secreting (only IL-17 expressing) Th17 cells. © 2011 The Society for In Vitro Biology.
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