24-Well Cell Invasion Assays, Basement Membrane

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Seed 1x10^6 cells	Incubate cells for 48 hours at 37ºC in 5% CO2 atmosphere.

Upstream tips
Seed 1x10^6 cells
Protocol tips
Incubate cells for 48 hours at 37ºC in 5% CO2 atmosphere.

Publication protocol

Influence on migration was conducted on MG-63 cells, pre-incubated in assay medium for 48 h adaption in 6-well plates (Greiner, Germany). A scratch wound was made by Ibidi culture inserts (μ-Dish 35 mm; Ibidi GmbH, Martinsried, Germany) following the instructors recommendations. When cell layers reached confluence, the culture insert was removed and cells were treated with VSM (25–50 μg/ml) extract or control (vehicle, DMSO). Gap closure was analyzed as described previously [50]. Cell invasion assay was performed with the CytoSelect™ 24 –Well Cell Invasion Assay (Basement Membrane, Fluorometric Format) from Cell Biolabs, Inc., CA, USA. Briefly, 1x106 MG-63 cells with the plant extracts were seeded in the membrane insert for 48 h. Fluorescence of invaded cells was counted with a plate reader at 480/520 nm.

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