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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

Biological characterization of a granulomonopoietic enhancing activity derived from cultured human lipid-containing macrophages.

We describe the biologic characteristics of an activity produced by human monocyte-derived lipid-containing cells (MDLCCs) that enhances the colony-forming capacity of granulocyte-macrophage progenitors (CFU-GM). Medium conditioned by well-developed MDLCCs (at day 21 to day 28 of cultivation) was added to bone marrow cultures containing GCT cell line-conditioned medium (GCT-CM) or other material as a source of granulocyte-macrophage colony-stimulating factors (GM-CSFs). MDLCC-conditioned medium (CM) had no detectable granulocyte-macrophage colony-stimulating activity (GM-CSA), but it contained an activity that enhanced the colony number in both day 7 and day 14 CFU-GM cultures. Dose-response curves for GCT-CM in the presence of MDLCC-CM demonstrated that this enhancing effect occurred at concentrations of GM-CSFs that stimulate maximal CFU-GM growth. This enhancing effect was seen with both granulocytic and monocytic progenitor cells. It was titratible and required the continuous presence of MDLCC-CM from initiation of culture. No enhancement was noted when MDLCC-CM was added 48 hours after plating. The enhancement still occurred when marrow cells were first incubated with MDLCC-CM and GCT-CM was added at later times. Neither the enhancing activity nor its production was dependent on horse serum contained in MDLCC culture medium. The enhancing effect was also seen when other sources of GM-CSA were used: medium conditioned by 5637 cell line, phytohemagglutinin-stimulated lymphocytes (PHAL), or placenta tissue. Furthermore, this enhancing activity appeared to be specific for CFU-GM. Addition of MDLCC-CM to mixed and erythroid cultures, stimulated by suboptimal and optimal concentrations of PHAL-CM did not modify the number of mixed colonies or erythroid bursts. This granulomonopoietic enhancing activity contained in MDLCC-CM was heat stable (56 degrees C and 75 degrees C for 30 minutes) and nondialyzable (3,500 and 14,000 molecular weight cut off tubing). Its production was increased by treating MDLCC with lipopolysaccharide (5 micrograms/mL) or zymosan (60 micrograms/mL) and inhibited by lactoferrin (10(-7) mol/L). The production of a granulomonopoietic enhancing activity by MDLCCs represents the demonstration of another positive feedback regulator of myelopoiesis involving the monocyte-macrophage system.[1]


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