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Saggi cellulari

High throughput assays for migration, invasion and more

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Documentation

Riferimenti bibliografici

Panoramica dei riferimenti bibliograficiApplicazione
Extracellular HIV-1 Nef increases migration of monocytes
Michael H. Lehmanna, Stefan Walterc, Loyda Ylisastiguid, Frank Striebelc, Vladimir Ovode, Matthias Geyerf, Jean Claude Gluckmand and Volker Erflea
Experimental Cell Research, Volume 312, Issue 18 , 1 November 2006, Pages 3659-3668  2005

High throughput in vivo screening for bone anabolic compounds with zebrafish
Angeleen Fleming, Masahiko Sato, Paul Goldsmith
Journal of Biomolecular Screening 10(8); 2005, 823-831  2004

Screening Assay for the Identification of Deoxyhypusine Synthase Inhibitors
Marc-Nicola Somner, Dorian Bevec, et al.
Journal of Biomolecular Screening, 2004, 9(5):434-438  2004

Cell Based Assays
A cell-based assay for screening the uridine 5'-diphosphate-glucuronosyltransferase 1A inhibitory potential of new chemical entities.
Jaime Padros, Donald Chun, Liangfu Chen, Pierre Rigolli, Themis Flarakos and Malle Jurima-Romet; Analytical Biochemistry 320 (2): 310-312
Analytical Biochemistry 320 (2): 310-312  2003

Cell Culture
Functional characterization of Glu298Asp mutant human endothelial nitric oxide synthase purified from a yeast expression system.
Regina Golser, Antonius C. F. Gorren, Bernd Mayer and Kurt Schmidt
Nitric Oxide 8 (1): 7-14  2003

Cell Based Assays

FAQ

DomandaRisposta
What is the well depth and maximum volume capacity of a MultiScreen plate?The well depth of a 96 well MultiScreen plate is 1.245 cm. The well depth for a 384 well MultiScreen plate is 1.2 cm. The maximum working volume of a 96 well plate is 300 ul. The maximum working volume for a 384 well plate is 100 ul.
Assay Terminology commonly used with reference to MultiScreen-MIC platesChemotaxis: Directed movement of cells in response to a concentration gradient.

Migration: Random movement of cells.

Invasion: The movement of cells across a barrier (e.g. extracellular matrix) in response to a concentration gradient.

Transmigration: The movement of cells across another cell barrier in response to a concentration gradient.

Angiogenesis: Tubule formation exhibited by HUVEC (Human Umbilical Vein Endothelial Cells) in response to cell stimulating factors.

Co-culture: When you have more than one cell type in your assay it is called co-culture. These assays are typically run to assess cell-cell interactions and factors secreted by one cell influencing the behavior of the other cell.
What is the MultiScreenTM-MIC plate?It is a 96 well sterile single use device that allows you to perform a variety of assays such as migration, invasion, chemotaxis, angiogenesis, transendothelial migration and co-culture type of assays. It has a filter plate* with 96 wells fitted with membrane. These wells serve as upper wells in your assay. It has a 96 well tear drop receiver plate. These wells serve as the lower wells in your assay. The membrane is the separation component between the two wells. These devices are available in a variety of pore sizes (3, 5 and 8 mm pore sizes available as cataloged products. 1, 2 and 12 mm pore sizes available by special order) in an uncoated format. The plates are available in packs of 10 with each device individually wrapped in a blister pack to ensure sterility. The plates are tissue culture treated (using a proprietary methodology) to support use with suspension and adherent cells.

*There is no underdrain on these filter plates, as the application does not require use of vacuum.
What type of assays can I perform on the MultiScreen-MIC plates?he MultiScreenTM-MIC plates can be used for a variety of assays such as migration, invasion, chemotaxis, angiogenesis, transendothelial migration and co-culture type of assays.
What type of screening can I perform with the MultiScreenTM-MIC plates?Some examples of screening that can be done on MIC plates are compound library screening, protein library screening or antibody library screening. You can measure the effect of the compound, protein or antibodies on cell behavior in functional assays such as migration, invasion, chemotaxis, angiogenesis, transendothelial migration and co-culture type of assays.
What pore size should I select when using the MultiScreenTM-MIC plate for my assay?Pore size determination depends totally on your cell type. A quick literature search will enable you to decide the best pore size for the particular cells you are using.

The following chart illustrates pore size choice for cell lines used in our laboratory and by some of our customers for these assays.
Cell NameCell TypePore sizes typically usedAssays typically performed
MDA-MB-231Invasive Breast cancer cell line (human)5 or 8 µm5 or 8 µm used in chemotaxis or invasion assay
MCF7Non-invasive breast cancer cell line (human) 5 or 8 µm5 or 8 µm used in chemotaxis or invasion assay
HT1080Invasive fibrosarcoma cell line (human)5 or 8 µm 5 or 8 µm used in chemotaxis or invasion assay
NIH3T3Non-invasive fibroblast cell line (mouse)5 or 8 µm5 or 8 µm used in chemotaxis or invasion assay
HUVEC (Human vein umbilical vein endothelial cells)Endothelial cells3 or 5 or 8 µm3 or 5 or 8 µm in chemotaxis, invasion, angiogenesis or transendothelial migration assays
HMVEC/HMEC(Human dermal microvascular endothelial cell)Endothelial cells5 or 8 µm5 or 8 µm in chemotaxis, invasion, angiogenesis or transendothelial migration assays
PMNPolymorphonuclear neutrophils1 or 3 µm1 or 3 µm in chemotaxis assays
-Primary Stromal cells8 µmNo information available
-Epithelial cells3 or 5 µm No information available
-Human coronary artery smooth muscle cells5 µmNo information available
Hepatic stellate cellsMyofibroblast 5 µmNo infor
What is the pore density in the MultiScreenTM-MIC plates?Pore size Nominal pore density per cm2 3 mm MIC 2 X 106 5 mm MIC 4 X 105 8 mm MIC 1 X 105
What is the surface area of membrane in the MultiScreenTM-MIC plates?The surface area of membrane in the MultiScreenTM-MIC is 0.3 cm2
What cell density should I use in my assay when using the MultiScreenTM-MIC plates?Every cell type calls for a different cell density. Before you set up your actual assay, you must run standardization experiments testing a range of cell densities and assess which cell concentration gives you the optimal results. You can adjust the cell: pore ratio to optimize your signal to background values. Pore densities for the different pore sizes are listed below.

We have used invasive and non-invasive breast cancer cell lines (MDAMB231 and MCF7), epithelial cell line (HT1080) and fibroblast cell lines (NIH3T3) in our MIC plates. We typically use 50,000 cells/well.

Pore sizePore density per cm
3 mm MIC2 X 106
5 mm MIC4 X 105
8 mm MIC1 X 105
What volumes should I use in my assay when using the MultiScreen-MIC plates?We typically use 50 µl in the top well and 150 µl in the bottom well.