Cell migration / Invasion cell type - HUVEC

Cell Invasion or Cell Migration assays are technically challenging to set up as the gradient between the two compartments equilibrates in time during the assay. It is also problematic to view cells and for cells to migrate through a non-physiologic polycarbonate or polypropylene filter. Care must be taken while loading the well with cells to form a single cell suspension. Precaution must be taken while trypsinization (under-trypsinization can lead to cell clumping while over-trypsinization could strip off adhesion molecules necessary for migration). This leads to difficulty in getting significant results, when only small numbers of cells cross the filter or when the distribution and/or staining of the cells is uneven.

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Found 3 matching solutions for this experiment

Protocol tips
Incubate cells for 30 min at 37 °C

Lower chamber is with 1% serum

Both chambers were assembled and incubated for 4 h
Upstream tips
Seed 2×105 cells in the top chamber
Protocol tips
Cover plate and incubate for 24 to 36 h hours at 37°C in a CO2 incubator (4-6% CO2).
Protocol tips
Incubate for 24 hours in a cell culture incubator
Downstream tips
Read fluorescence with a fluorescence plate reader at 480 nm/520 nm.
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