Tubes - BD

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Bd Biosciences - Discovery Labware BD HUVEC-2 cells assayed in the BD BioCoat Angiogenesis System: Endothelial Cell Migration (96-Multiwell format) in response to increasing concentrations of VEGF . Samples were incubated for 22 hours . Cells were labeled post-migration with Calcein AM and measured by detecting the fluorescence of cells that migrated through the fibronectin-coated BD FluoroBlok membrane with the Victor 2 plate reader (Perkin Elmer) at 485 nm emission . Data represents the mean of four inserts ± SD . TIP Fold increase over control (mean ± SD) 4 3 2 1 0 Control 1 5 10 VEGF (ng/ml) description Qty. cat. no. Bd Biocoat angiogenesis system: endothelial cell migration 24-Multiwell Insert System 1 354143 24-Multiwell Insert System 5 354144 96-Multiwell Insert System 1 354147 96-Multiwell Insert System 5 354148 Remember to order the respective BD Falcon FluoroBlok Multiwell Insert System as control plates when purchasing the BD BioCoat Angiogenesis System: Endothelial Cell Migration . RELATED PRODUCTS More BD BioCoat Angiogenesis Systems . . . . . . . . . . . . . . . . . . . . . . . . . . this chapter BD HUVEC-2 Cells . . . . . . . . . . . . . . . . . . . . 154 BD BioCoat Endothelial Cell Growth Environment . . . . . . . . . . . . . . . . . . 68 BD Vascular Endothelial Growth Factor . . . . . . . . . . . . . . . . . . . . . . . . . . 194 drug discovery and development 7 insert systems bdbiosciences.com | 151
7 insert systems drug discovery and development Bd Biosciences - Discovery Labware Bd Biocoat angiogenesis system: endothelial cell tube Formation • Save time and improve reproducibility with optimized system for screening compounds that modulate endothelial cell tubulogenesis Endothelial cells are capable of differentiating in vitro to form capillary-like structures (also referred to as tubes) 1,2 . Cross section analyses of tubes formed in extracellular matrices such as laminin 3 , collagen 2 , fibrin, and BD Matrigel Matrix 3 revealed the presence of a lumen surrounded by cells. When BD Matrigel Matrix is used to promote endothelial cell tube formation, the lumen forming cells interact with each other via interdigitating junctional complexes 3 . This property is indicative of in vivo-like capillary formation. Since BD Matrigel Matrix has been found to induce endothelial cell tubulogenesis and lumen formation in a timeframe of 18 hours, this matrix is suitable for high throughput assays that target this critical angiogenesis pathway. The BD BioCoat Angiogenesis System: Endothelial Cell Tube Formation is an in vitro assay system composed of a BD Falcon 96-well black plate with clear bottom uniformly coated with BD Matrigel Matrix. To insure reproducibility when using this assay system, different preparations of BD Matrigel Matrix are screened for the ability to promote optimal tube formation under standardized conditions. In addition to the coating material, the manufacturing process has been optimized to the highest standards. Assay performance is further enhanced by the inclusion of our specially treated 96-well microplate, which has specific surface properties that assure even coating and minimize meniscus formation. 152 | bdbiosciences.com Confocal Imaging of BD HUVEC-2 Cell BD HUVEC-2 cells (Cat . No . 354151) were assayed using the BD BioCoat Angiogenesis System: Endothelial Cell Tube Formation (Cat . No . 354149) . Cells were stained using the fluorogenic esterase substrate Calcein AM (Molecular Probes C1430) . Confocal images were captured using the BD Pathway Bioimager equipped with a 4 X Olympus objective . The 96-well format allows for increased productivity in this traditionally low throughput assay. Throughput can be further augmented through the use of automated imaging instrumentation to obtain optimal microscopic images for rapid quantification of tube formation. A number of human endothelial cell types have been shown to form tubes in the BD BioCoat Angiogenesis System: Endothelial Cell Tube Formation. Studies have demonstrated that endothelial cell tubulogenesis is inhibited by antagonists to integrin subunits ·5ß1 4 and ß1 5 and the antibiotic fumagillin 6 . Using the BD BioCoat Tube Formation System, we have shown that suramin and antibodies directed against ß1-integrin inhibit endothelial cell tube formation in vitro. REFERENCES 1. Folkman, J. and Haudenschild, C., Nature 288:551 (1980). 2. Montesano, R ., Orci, L, and Vassalli, J., Cell. Biol. 97:1648 (1983). 3. Kubota, Y., Kleinman, H.K., Martin G.R. and Lawley, T.J., J. Cell Biol. 107:1589 (1988). 4. Kim, S., Bell, K., Mousa, S.A., and Varner, J.A., Am. J. Pathol. 156:1345 (2000). 5. Kubota, Y., Kawa, Y., and Mizoguchi, M., J. Dermatol. Sci. 12:36 (1996). 6. Ingber, D., et al., Nature 348:555 (1990).
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204 Bioimaging SyStemS BD Bioscienc
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9 Pathway Bioimager Bioimaging SySt
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9 aCtone gPCR Screening assay Bioim
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10 Alphabetical Index AlphAbetIcAl
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10 numerical Index nUMeRIcAl Index
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BD Biosciences Worldwide Discovery
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Tubes - BD

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