monitor cell growth - (Sep/25/2006 )
I want to monitor cell growth of a certain cells on petri dish. The cells are adherent and are passaged by scraping using a cell scrapper.
My plan is to first dilute a suspension of cells to about 10^6 cell/mL and culture them on 60mm petri dish. Then after 24 hours I'll pour off the media and put in 5mL of fresh media, scrap the cell off and count the number again.
Does that sound reasonable?
your plan looks fine, just few reminders : 1) duplicate or triplet your sample to get an average mean of cell number following time course (seed cells in 24-well plate will be easier than dish.) 2) tripan blue staining when count cells - I assume you don't want to count the dead cells ?! 3) if your cells express certain distinguishable surface marker, then stain for it and FACS it to get more accurate cell population. 4) CFSE stain helps to monitor cell differentiation using FACS (the stain will be diluted from parent cells to daughter cells, therefore you will see less and less fluorescence).