Jurkat Cell Adhesion - Techniques to make Jurkat Cells to adhere to plastic surfaces (May/18/2008 )
I am trying to study calcium flux in Jurkat cells using fluorescence. In order for the fluorescence microscopes camera to detect the region of interest after the cells are stimulated (i.e. addition of agonist) I need the cells to stick to the plastic 96-well plate.
I have tried using fibronectin. This works as an adherent substratum for the Jurkat cells but unfortunately also triggers a spontaneous calcium current via VLA receptor cross-linking. Not very convenient .
I have unsuccessfully attempted to use Poly-D-lysine (MW < 70 KDa) at a concentration of 1 mg/mL but the cells don't stick to the plastic plate. The Poly-D-lysine works for all of my other cell lines (F11, HEK). I have noticed that in the literature that labs only seem to be using glass plates when trying to get Jurkat cells to adhere to a surface. I feel like there is something I'm missing i.e. plastic isn't a suitable surface perhaps for Poly-D-lysine? Not sure.
In the meantime I'm going to try a higher molecular weight Poly-D-lysine (MW < 100 KDa) and Poly-L-Ornithine. Not sure what to try after that. Perhaps Laminin or collagen?
Any suggestions would be greatly appreciated.
I have seen a colleague coating multi chamber slides with Poly-L-Lysine, before seeding Jurkat. I don't know, is there a difference between D and L ?
Poly-D-lysine is more resistant to enzymatic degredation. Making it a more reliable adherent. Other than that not sure what the differences in adherent properties would be between the two.