I need to do migratation assay with pc 9 cell. I am not getting consistent results as I used different cell numbers in the assay.
If anyone can help me if cell density in the assay does impact in the output of effective result.
hi,
Cell density play important role in cells migration. Its gives some kind of force to cells to migrate. So keep same number of cells every time.
For doing cell migration, the density is very important. In the condition with low density of cells, cells will migrate faster than in the high density of cells. I often use 0.25x10^4 cells per well (96 wells plate) for NIH3T3 cell line. I think every cell line we have to optimize it ~
cell density is very vital for cell migration. when doing wound healing assay (in vitro scratch assay), the cells need to become confluent to in order to promote proper cell migration. if they are sparse they would move to fill the space in between them, however, for confluent cells they would move to the created wounded area. The same goes for Transwell migration assay. the cells need to be seeded at high density to increase their rate of migration onto the membrane. i have attached two links. the first link is a nature protocol describing how to perform wound healing cell migration assay. for the second link, it contains a software called TScratch that would automatically calculate the rate of migration for you. It is automated and therefore reduces error in estimating cell migration.
cheers bro!!!!!!!!!!!!!!!
http://www.nature.com/nprot/journal/v2/n2/pdf/nprot.2007.30.pdf
http://www.biotechniques.com/multimedia/archive/00045/BTN_A_000113083_O_45150a.pdf
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