Dear Researchers,
Hi,
How can I use clinical sample like plasma, which is tested for viral infection, in cell culture propagation?. Basically, I do know the sample is positive for virus and what kind/type of cell is required, but I'm not certain of propagation procedure and its initial preparation steps. I'm looking for your experience for such expriment.
Regards,
Alireza
hi,
there are different types of mammalian cell lines are there which cab be used for virus propagation. Type of cell depends upon which virus you want to propagate. so first tell me which virus you want to propagate?
for plasma cells, you just do two or three cycle of freeze thaw than take the supernatant for infecting the cells.
Dear Laxmi,
Hi,
I want to infect HFF cell line by a clinical sample positive for HCMV virus. By freeze and thaw and centrifugation I will get the virus in the suppernatant?
Is furthere decontamination is neccesary or pen/strep will do?
yes by freeze thawing you will get the virus in the supernatant.
peniciline/strep will be sufficient.
all the best
Just remember that repeated freezing and thawing often destroys virus particles too so it maybe worth holding onto some of the original sample. Also, centrifuge before inoculating cultures will also help chances of contamination.
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