Cell migration / Invasion cell type - SaOS-2

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
- Seed viable cells at 2×105/200μL after transfection in a serum free medium on theupper side of the well.

- Add complete medium on the lower side.
Protocol tips
- Treat upper chamber with Matrigel matrix membrane after 48h of culture.
Downstream tips
- Stain with 0.2% w/v crystal violet before counting cells.
Protocol tips
Add 10% fetal bovine serum) to the lower chamber.

Cover plate and incubate for 24 - 72 hours at 37°C in a CO2 incubator (4-6% CO2).
Upstream tips
Seed 4 × 10^4 cells
Protocol tips
Incubate cells at 37 °C for 24 hours
Downstream tips
Read plate at 485/520 nm filter set.
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