Effect of CXCR1 and CXCR2 Receptors Inhibition on Megakaryocyting Differentitation of Umbilical Cord Blood CD 133+ Cells (Pages: 0-0)


Khalaf Adeli E *, Kheirandish M , Abolghasemi H , Ebtekar M , Siadat S.D , Noroozi Aghideh A , Haghbin S ,

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Objective: Most studies have reported that some CXC chemokines including NAP-2, IL-8 and PF4 which expressed by megakaryocytes in megakaryocytopoiesis and affect cell expansion and differentiation of hematopoietic stem cells. Previous studies have shown that these chemokines inhibit megakaryocytopoiesis .The receptors for NAP-2 and IL8 and most likely PF4 were reported to be CXCR1 and CXCR2. The aim of this study was to investigate the effect of inhibition of CXCR1 and CXCR2 on differentiation of umbilical cord blood (UCB) CD133+cells into megakaryocyte progenitor cells. Materials and Methods: Umbilical cord blood CD133+ cells were separated by magnetic cell sorting method. CD 133+ cells were placed immediately after selection, in a serum free medium supplemented with IL3, IL 6, TPO and stem cell factor (SCF) as well as 5% CO2 for 12 days as a control cells. To investigate the effect of receptor inhibition, the CD133+ cells were cultured under a same condition with cocktail cytokine and divided into three groups: in the first group only CXCR1 was blocked , in the second group only CXCR2 blocked by neutralizing monoclonal antibody, in the third group both receptors were blocked by Anti CXCR1/CXCR2 neutralizing monoclonal antibodies. Expression of the CD 41 and CD61 antigen as megakaryocyte progenitor cell markers were evaluated by flow cytometry. Results: The results showed that inhibition of CXCR1 and CXCR2 receptors together caused an increase in CD61 expression on days 7 and 12 in comparison to cells treated with IL3, IL 6, TPO and SCF (p