Cell migration / Invasion cell type - TPC1

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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Cultrex® Collagen IV Cell Invasion Assay

Trevigen

Upstream tips	Protocol tips	Downstream tips
	We used Cultrex 24-well collagen IV cell invasion assay kit (Trevigen, Gaithersburg, MD, USA). Transwell cell culture inserts with an 8-μm pore membrane were coated with collagen type IV. Cells (2 × 105 cells/insert) were seeded onto the upper chamber without FBS, and 500 μL of RPMI 1640 containing 10% FBS was loaded onto the bottom chamber. After 48 h of incubation, the cell dissociation solution was added onto the bottom chamber and incubated 1 h.

Protocol tips
We used Cultrex 24-well collagen IV cell invasion assay kit (Trevigen, Gaithersburg, MD, USA). Transwell cell culture inserts with an 8-μm pore membrane were coated with collagen type IV. Cells (2 × 105 cells/insert) were seeded onto the upper chamber without FBS, and 500 μL of RPMI 1640 containing 10% FBS was loaded onto the bottom chamber. After 48 h of incubation, the cell dissociation solution was added onto the bottom chamber and incubated 1 h.

Protocol tips

We used Cultrex 24-well collagen IV cell invasion assay kit (Trevigen, Gaithersburg, MD, USA). Transwell cell culture inserts with an 8-μm pore membrane were coated with collagen type IV. Cells (2 × 105 cells/insert) were seeded onto the upper chamber without FBS, and 500 μL of RPMI 1640 containing 10% FBS was loaded onto the bottom chamber. After 48 h of incubation, the cell dissociation solution was added onto the bottom chamber and incubated 1 h.

Manufacturer protocol Publication protocol 1 Relevant paper

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