in the first experiment, we incubated individual purified proteins (e.g., FN, VN, laminin, and collagen) diluted in PBS solution on nano structures for 24 hrs, followed by deposition and incubation of Vero cells in OptiPro medium for 24 hrs.
in the second experiment, we directly deposited the Vero cells in a solution of OptiPro+individual proteins (VN, FN, Laminin, and Collagen) on teh same nano-structures for 24 hrs.
Results indicate that Vero cells alignment and population from the first experiment were higher.
Any thought for why?
Consider this: In expt 1 the only sites available for your VN, FN, LN, & COL substrates to bind to your nanostructured material are those of the material. In expt 2, you include additional sites present on the Vero's. If the concentrations of VN, FN, LN, & COL are limiting (i.e. not saturating) then the Veros will act as a sink and reduce the effective availability of molecules binding to the material. Simple Michaelis-Menton kinetics - you have altered the equilibrium equation!
To interpret these expts you need to understand how cells interact with dispersed versus polymerised ligands, concepts of equilibrium and fluid dynamics.
Consider also: the interactions between VN, FN, LN, & COL and the material are distinct and inequivalent to the interactions these species have with the Veros.
Lastly, I assume that your Veros are grown in media with FCS. What are the major species present in FCS? FN, VN, some LN and some COL!! How well did reduce contamination from the FCS? If it was anything less than 6 washes with
Thank you David. the medium in which Vero cells were incubated originally was OptiPro,serum free, low protein content culture media less than 6um/ml.
- Badges
- Science topic
- Similar topics
- Chemistry
- Nanotechnology
- Nano