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1.Suspension cell preparation:Before adenovirus stabilization of the transfected cell line, please ensure that the target cells are in good condition and free of mycoplasma contamination. A cell confluence rate of 40% to 60% is appropriate for adenovirus infection.
2.Determination of optimal MOI of infected cells:MOI (Multiplicity of infection) is the number of viruses infecting each cell.
Note: The optimal MOI of adenovirus varies for different types of cells from different sources. For suspension cells, the MOI of adenovirus is generally graded in increments of 30, 100, 300, 1000, 2000. Adenovirus MOI for suspension cells is more demanding than that for adherent cells, and the MOI gradient is recommended to be performed in 96-well plates to save virus and cells.
3.Conversion of MOI to virus volume
Number of cells at the time of infection (generally calculated as the number of cells counted in the passages x 2) x MOI = number of viruses; then the number of viral volumes added = number of viruses/viral titer
Ads infection of suspension cells:
Viral: Ads-GFP,1×10¹º PFU/ml
Cell of infection: OCI-LY1、K562、HL-60、L2110
Detection Methods: Detection by Fluorescence microscopy for 72 h after infection
Ads infection of mouse primary CD4+ T cells:
Cell : mouse primary CD4+ T cells
Mode of Stimulation: CD3/CD28 antibody and IL-2 activation for 48 hours
Viral:Ads-GFP, 1×1010 PFU/ml
Ads infection of human primary CD4+ T cells:
Cell : human primary CD4+ T cells
Mode of Stimulation: CD3/CD28 antibody and IL-2 activation for 48 hours
Viral:Ads-GFP, 1×1010 PFU/ml
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