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ARTICLE
Year : 2011  |  Volume : 1  |  Issue : 1  |  Page : 38-49

Immunophenotypic Study of the Cord Blood CD34+ Progenitor Stem Cells-Derived Dendritic Cells


1 National Centre of Hematology research and treatment-Al-Mustansiriyah University
2 Al-Nahrain University, college of medicine, Dept. of Microbiology
3 Al-Nahrain University, college of medicine, medical research unit

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Background: Dendritic cells (DCs) are bone marrow-derived cells of both lymphoid and myeloid stem cell origin that populate all lymphoid organs including the thymus, spleen, and lymph nodes, as well as nearly all no lymphoid tissues and organs. Although DCs are a moderately diverse set of cells, they all have potent antigen-presenting capacity for stimulating naive, memory, and effector T cells . DCs are receiving increasing scientific and clinical interest due to their key role in anti-cancer host responses and potential use as biological adjuvants in tumour vaccines, as well as their involvement in the immunobiology of tolerance and autoimmunity . Objectives: To generate the dendritic cells (DCs) in vitro from purified cord blood stems progenitor cells and detects the growth curve of cells and analyze the DC in vitro differentiation pathway. Also to investigate the immunophenotypic CD markers morphology of the surface DC cells and to describe the effect of different growth factors on their expression. Materials & Methods: For isolation and establishment in tissue culture of human DC, the cord blood was obtained from placenta of newly vaginaly delivered women, who were admitted in Al Kadhmyia Teaching Hospital. The cells were cultured in complete RPMI growth medium supplemented with growth factors GM-CSF+IL-4 for seven days then the key surface CD markers were detected by using the immunocytochemistry technique. Results: The use of the CD34 monoclonal antibody in combination with CD45 monoclonal antibody has increased the opportunity of obtaining a reasonable amount of purified stem cells. Furthermore, the growth factors (GM-CSF+IL-4) that were supplemented to the complete medium played an important role in the differentiation of the CD34+ stem cells toward the DC cells. The growth factors GM-CSF and IL-4 when used in combination had made the difference and the rhythm of differentiation ,the count of cells, resolution of results, shape and size of cells, and the state of DC cells better than using each of them alone. The IL-4 when used in combination with GM-CSF act as inhibitory factor for the granulocyte and cells other than DC and at the same time has the capability to keep the DC cells in immature state. The kinetics of DC development in cord blood cultures was also determined. These results indicated that, the optimal culture of the cord blood-derived DC was 7-9 days. By using the immunocytochemistry technique, the key CD markers of DC cells has been revealed since the day 6 of culture. The immature DC cells were characterised with the positive expression of CD1a, CD11b and CD11c while the CD14 was negative. Conclusion: Isolation, purification and mobilization of stem cells obtained were reliable and confirmatory, and indicate that the positive differentiation of the cord blood CD34+ stem cells toward the DC cells was achieved.


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