In my experiment to study the migration of microglial cells, I employ scratch assay/wound healing assay. I wonder if it is compulsory to use Mitomycin C to ensure the scratch covered by the cell is due to migration, not because of proliferation.
My incubation time is 24 hrs and the doubling time of the BV2 cell line seems to be around ~35h. Any suggestion?
thanks in advance,
Hilmi
Hello Hilmi Jaufer T a
For longer term wound healing assays (> 24 hr.), you will require mitosis inhibitors such as mitomycin C treatment to eliminate the contribution of cell proliferation to gap closure, allowing the assay to specifically assess migration of the given cell population.
However, in your case, with incubation time of 24 hours and the doubling time of BV2 cell line being around ~35 hours, it may not be necessary to use mitomycin C.
Best.
Even when long times are required, contribution of proliferation is close to zero. You realize that when you make a time-lapse to monitor migration. At the leading edge there is no proliferation at all. A cell either proliferate or migrate, those are, obviously, exclusive cellular processes
- Badges
- Science topic