Search for: [Abstract = "The aim of this study was to design the methodology of expansion and differentiation of CD34\+ cells to monocytes\/macrophages. The present study shows that optimal medium for expansion of CD34\+ cells was X\-VIVO 10 supplemented with FBS, SCF, IL\-3, FLT\-3L, TPO and 3\-4 days culture. For differentiation of CD34\+ cells to monocytes\/macrophages was IMDM supplemented with FBS, M\-CSF, SCF, IL\-3, FLT\-3L and culture for 7\-10 days. Using this protocol it was possible to obtain 1000 \- fold increase in number of expanded cells, of which 40\-60% were CD14\+, i.e. 600 \- fold increase in the number of CD14\+ cells. The presence of two subpopulations of monocytes\: CD14\+CD16\- and CD14\+\+CD16\+, which occurred at the ratio of about 2\:1 was observed. They are clearly distinct from known main subpopulations of blood monocytes CD14\+CD16\+ and CD14\+\+. Supplementation of cultures with VD3 increased the percentage of CD14\+CD16\- and decreased the proportion of CD14\+\+CD16\+. Monocytes\/macrophages, mainly CD14\+\+\/16\+ subpopulation secreted IL\-12, but not TNF and IL\-10, induced allo\-MLR \(mainly CD14\+\+\/16\+ subpopulation\), fagocytosed S.aureus \(mainly CD14\+\/16\- subpopulation\), but did not produce intracellular O2\-. Hence, this study shows that it is possible to generate from CD34\+ haematopoietic progenitors two subpopulations of human monocytes. They are clearly distinct from those present in the peripheral blood implicating the existence of novel monocyte subsets with distinct biological function that represent different stages in monocyte differentiation."]

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