again thanks!!

i've done some preliminary tests, what i suspected was the period of keratinocyte adherent might be due to some factors. around the results i have got was the seeding density. if i seed too less cells, cell could adhere but need long period to get 60% of confluency and of course some cells were proliferated and DIFFERENTIATED. this is worse!

if too much cells, medium will need to change everyday, but 80% confluency can be achieved within 4 days. donor variability also a factor in this aspect as well.

what do u think about the seeding density??

thanks for comments!!

hi
you must pay attention to the time cells need to differenciate. if your cells need 5 days to differenciate, i would say that three days is the max time for your experiment. But if time of differenciation is greater no problem fo doing your experiment on 4 days!
I think that 60% confluency is good. For two reasons. 1 cells are relatively close to make good divisions (but not too close) and 2 if you seed at more than 60% and due to the fact your treatment is an increaser of division frequency (PDT decreases) more than 60% may result in too much confluency in treated cells.
Enjoy your experiment. Good luck.

Fred.

thanks,fred!!

u are right. i'll try through out the passage 0 to 2 with treatment, to observe the differences of PDT among passage 0,1 and 2. what i guess was the passage 2 will be cleaner than the other two. thus, will be giving out the more accurate answer. what do u think??

thanks!!

Anyone here would suggest any better idea,websites or journals that there were people had done it before, at least regarded.


SvenCondon
gas rc planes

those units which adhered might be less than 500 000 units, let state 50 000 units only get adhered, and other ones might be adhered 2 or 3 days later...how manage u deal with it?? one time u trypsinize your units for number assessment, manage u re culture your calculated units into the plates