Q: How is the gel released from the well?
A: After a two day culture, a sterile spatula is carefully inserted between the plate wall and the gel. The spatula is then slowly moved around the wall in a circle to completely release the gel. We use a small spatula with a thin, flat, edge and is the same spatula used for weighing out chemicals in milligram quantities. Prior to releasing the gel, the spatula should be sterilized with 70% ethanol.
Q: For time-course measurements, should the gel be removed from the well?
A: It is recommended to keep the gel in the 24-well plate when measuring the length of the gel over time. The gel is very soft and would be difficult to measure if removed from the plate.
Q: Does this assay only work on fibroblasts?
A: The collagen-based contraction kit is not just for fibroblast, it can be used for any cell types as long as the cells can be cultured in 3D collagen. Besides fibroblasts, people also study endothelial cell, muscle cell and heart cell contraction using the collagen model.
Q: Do you have a protocol for using this kit to culture cells in a 3D collagen matrix?
A: To use our Contraction Assay Kit (CBA-201) for culturing cells in a 3D collagen matrix, the assay protocol can be followed up to step 4. It is the same protocol but without the step for releasing the gel. Cells can be visualized using an inverted microscope.
Q: Can I recover cells from this assay?
A: We haven’t tried to recover cells from this assay and therefore we don’t have a protocol for this, but it is possible to do. The cells are in bovine type I collagen gel, which can be digested with a collagenase I incubation. The units required for digestion will require optimization.