Dear experts, I need to standardise and select the most appropriate cell seeding density for my stem cells to see the effcet of drug and treatment on cell viability and gene expression. Please enlighten how should I proceed and which plate format is best to carry out the experiment?
Thanks for your attention
Dear Priya, it depends if you want to see your cells proliferate or if you just want to see if they survive. According to that, you may seed at low density around 10000 cells/cm2 or at confluency around 50000 cells a cm2.
THank you very much for the information. I am planningto look into both survival and proliferation of stem cell after treatment with drug.
Hello
Agree with Benjamin Fournier
seed at low density around 10000 cells/cm2 or at confluency around 50000 cells a cm2.
Look at paper here will help you
Good Luck
http://www.hindawi.com/journals/sci/2012/920581/
http://www.sciencedirect.com/science/article/pii/S1873506110000978
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2289788/
http://www.stempeutics.com/pdf/24.pdf
http://www.ecmjournal.org/journal/papers/vol021/pdf/v021a37.pdf
It also depends on which assay you are using. If you're designing your own, the previous commenters are absolutely correct. If you are using, say, the Promega ApoTox Glo assay (https://www.promega.com/~/media/files/resources/protcards/apotox-glo%20triplex%20assay%20quick%20protocol.pdf), it is recommended you use
it depends on your assay meanwhile your media,, but normally if you are using media+10% FBS+5ng bfgfg, you cans seed only 3000 cell/cm2
Dear Priya
Seeding density will depend upon the duration of drug. How many cell cycles you expect your culture should face the drug treatment? Before conducted the drug administration you study the population doubling time of your cells in particular condition to get clear conclusive results. Your drug may be totally cytotoxic at different cell stages.
good luck
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