How to inactivate H1N1 virus in Cell culture supernatant ? - (Apr/05/2014 )
I am studying the effect of cytokines that are released from one H1N1 infected cell line (A549 cell) on miRNA expression from another contact non H1N1 infected THP-1 cell.
Of course the nice way is to go for separate coculture where the pore size is smaller that H1N1 virus and allow only Cytokine passage. Unfortunately, the smallest pore size is 0.4 µm which is larger than the H1N1 (0.3µm).
Now I think about getting the infected A549 cell supernatant and inactivate the H1N1 virus to leave only the cytokines in it. and transfere these supernataant to THP-1 cells and measure miRNA.
First question : With the aim of testing whether cytokine release from H1N1 infected A549 cells would affect miRNA expression of THP-1 cell, WHat is your opinion about this design ?
Second: How can inactivate H1N1 in cell supernatant ( by UV light for example )
Third, I know to use MDCK cells to test that the virus is efficiently inactivated. Is there is another methode to show effective inactivation of the virus ?
Could you dialyse out the cytokines and retain the H1N1 or the other way around?
I want to remove the virus particle from this supernatent to retain the cytokine only . By this, I will be sure that any miRNA expression observed upon addiding this supernatant to THP-1 cell is due to cytokine ( not the virus). Because simply I can check virus effect by just infect THP-1 cells with H1N1
You could possibly immunoprecipitate the viral particles, but I don't know if this would be very efficient.