Key Spec Table
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|Storage Conditions||Lyophilized: Stable for 2 years at -20°C from date of shipment. Rehydrated: Stable for 3 months at -20°C.|
|Reference overview||Application||Pub Med ID|
|Catecholamines up-regulate lipopolysaccharide-induced IL-6 production in human microvascular endothelial cells |
Gornikiewicz, A., et al
Faseb J, 14:1093-100 (2000) 2000
|The MRE11-NBS1-RAD50 pathway is perturbed in SV40 large T antigen-immortalized AT-1, AT-2 and HL-1 cardiomyocytes |
Lanson, N. A., Jr., et al
Nucleic Acids Res, 28:2882-92 (2000) 2000
|High-efficiency transient transfection of endothelial cells for functional analysis. |
Kovala, A T, et al.
FASEB J., 14: 2486-94 (2000) 2000
The definition of signaling pathways in endothelial cells has been hampered by the difficulty of transiently transfecting these cells with high efficiency. This investigation was undertaken to develop an efficient technique for the transfection of endothelial cells for functional analyses. Cells cotransfected with plasmid expressing green fluorescent protein (GFP) and the plasmid of interest were isolated by fluorescence-activated cell sorting (FACS) based on GFP expression. In the sorted cell population, a 2.5-fold enhancement in the number of cells expressing the gene of interest was observed, as confirmed by FACS analysis and Western blotting. Sorted cells retained functional properties, as demonstrated by chemotaxis to the agonist sphingosine 1-phosphate (SPP). To demonstrate the usefulness of this method for defining cellular signaling pathways, cells were cotransfected with plasmids encoding GFP and the carboxyl-terminal domain of the beta-adrenergic receptor kinase (beta ARKct), which inhibits signaling through the beta gamma dimer of heterotrimeric G-proteins. SPP-induced chemotaxis in sorted cells coexpressing beta ARKct was inhibited by 80%, demonstrating that chemotaxis was driven by a beta gamma-dependent pathway. However, no significant inhibition was observed in cells transfected with betaARKct but not enriched by sorting. Thus, we have developed a method for enriching transfected cells that allows the elucidation of crucial mechanisms of endothelial cell activation and function. This method should find wide applicability in studies designed to define pathways responsible for regulation of motility and other functions in these dynamic cells.
|Purine nucleotides induce regulated secretion of von Willebrand factor: involvement of cytosolic Ca2+ and cyclic adenosine monophosphate-dependent signaling in endothelial exocytosis |
Vischer, U. M. and Wollheim, C. B.
Blood, 91:118-27 (1998) 1998
|Suppression of p53 activity and p21WAF1/CIP1 expression by vascular cell integrin alphaVbeta3 during angiogenesis |
Stromblad, S., et al
J Clin Invest, 98:426-33 (1996) 1996
|Isolation, growth requirements, cloning, prostacyclin production and life-span of human adult endothelial cells in low serum culture medium. |
Hoshi, H and McKeehan, W L
In Vitro Cell. Dev. Biol., 22: 51-6 (1986) 1986
Endothelial cells from autopsy and biopsy specimens from a variety of adult human vascular tissue were harvested by collagenase treatment and gentle swabbing of the lumenal surface. Nutrient medium MCDB 107 containing a partially purified brain-derived growth factor (5 micrograms/ml), epidermal growth factor (10 ng/ml) and only 2% (v/v) fetal bovine serum supported clonal and long-term serial culture (17.6 to 26.1 cumulative population doublings) of endothelial cells from vena cava, thoracic aorta and tibial arteries at a 70% rate of success. Cumulative doublings of the cell population from eight cultures were inversely proportional to age of donor of the vascular tissue from which cells were isolated. Heparin had an enhancing effect on cell growth that varied with cell strain. Prostacyclin production of human adult endothelial cell cultures was stimulated by arachidonate and thrombin by 17 to 20 and 2 to 3-fold respectively. Endogenous and stimulated rates of prostacyclin production by human adult endothelial cells were 2 to 3 times that of human adult smooth muscle cells and 20 to 30 times that of human fibroblasts.
Progress in hemostasis and thrombosis, 7:167-82 (1984) 1984
|Endothelial cell growth supplement: a cell cloning factor that promotes the growth of monoclonal antibody producing hybridoma cells |
Pintus, C, et al
J Immunol Methods, 61:195-200 (1983) 1983
|An endothelial cell growth factor from bovine hypothalamus: identification and partial characterization |
Maciag, T, et al
Proc Natl Acad Sci USA, 76:5674-8 (1979) 1979