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|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.
|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).
|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.
|Incubate the Corning BioCoat™ Matrigel Invasion Chambers for 20 hours in a humidified tissue culture incubator, at 37o C, 5% CO2 atmosphere.
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