A research update
In a previous post, I talked about the difficulties I had getting cells to stick on the surface of my hydrogels. Here’s an update:
I’ve been trying to find the right concentration of RGD such that the cells will not only attach to the hydrogels, they will also proliferate at a rate comparable to what they’d do on a plain tissue culture plate. If the RGD concentration is too high, the cells will attach, but not proliferate. I tried an experiment using different concentrations from 100 μM (micromolar) to 1 mM (millimolar, ten times the 100 μM), and found I was getting the best results at about 500 μM.
However, I was still getting more cells falling off the hydrogel surface than staying on. I thought, if cells landed on the surface and attached, would they stay attached after movement? That way, I could move the hydrogel to a new well, anything that was on the surface would stay on the surface. Any cells on the bottom of the tissue culture plate would stay there. The well I would move it to would only have cells on the surface.
I tried this by moving hydrogels at specified time points after adding the cells-2, 4, 6, 8, and 24 hours. Unfortunately, cells still fell off the hydrogels after movement, no matter what time point I moved them.
After sharing these results with my PI, he recommended trying experiments with single-layer hydrogels instead of bilayers. I would be lying if I said I wasn’t disappointed. But as we know, projects not working as expected goes along with the territory.
I’m starting a new round of cell experiments next week. We’ll see how they work!