Cell migration / Invasion cell type - BxPC-3

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

Upstream tips
Add 300 µL of prewarmed serum free media to the interior of the inserts.

Allow this to rehydrate the ECM layer for 15-30 minutes at room temperature.
Protocol tips
Add 250 µL of prepared cell suspension from step 5 to each insert.

Add 500 µL of serum free media in the presence or absence of chemoattractant.

Cover plate and incubate for 24 - 72 hours at 37°C in a CO2 incubator (46% CO2).
Upstream tips
Add warm (37C) bicarbonate based culture medium to the interior of the inserts and bottom of wells. Allow to rehydrate for 2 hours in humidified tissue culture incubator, 37C, 5% CO2 atmosphere.
Protocol tips
Incubate the Corning BioCoat™ Matrigel Invasion Chambers for 20 hours in a humidified tissue culture incubator, at 37o C, 5% CO2 atmosphere.
Protocol tips
Incubate at 37 °C in a tissue culture incubator for 72 hours to promote spheroid formation.

Downstream tips
Incubate at 37 °C in a tissue culture incubator for 6 days, and photograph the spheroid in each well every 24 hours using the 4X objective
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