The Suspended cell specific adenovirus（Ads）developed by HANBIO has been modified for the cell membrane characteristics of suspension cells, and the virus infection system has also been optimized accordingly, which greatly improves the infection ability for suspension cells. It has no effect on the infection ability of adherent cells and can be used universally. The infection efficiency of HANBIO Ads is close to 100% for many suspension cell lines such as Jurkat, K562, etc., and can reach more than 60% for primary cells of blood origin, such as CD4+ T cells.
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.
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.
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:
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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