Preparation method for adenovirus p53-loaded dendritic cell vaccine

A preparation method for an adenovirus p53 (Ad-p53)-loaded dendritic cell (DC) vaccine, comprising the following steps:

• (1) collecting peripheral blood and centrifuging the same, separating blood cells and autologous plasma, collecting an upper plasma layer, inactivating the plasma layer and centrifuging the same again, collecting inactivated plasma supernatant as the autologous plasma for later use, diluting cells with the plasma layer removed with normal saline before being mixed well with a lymphocyte separation solution in a ratio of 2:1, adding a mixture into a centrifuge tube comprising the lymphocyte separation solution for centrifugation, and sucking a buffy coat to collect peripheral blood mononuclear cells (PBMC), and centrifuging and washing the PBMC again;
• (2) collecting CD14+ cells and CD14− cells by incubating and sorting the PBMC with CD14 magnetic beads, and performing an adherent culture of the CD14+;
• (3) discarding a supernatant from the CD14+ cells after completing the adherent culture in step (2), adding a DC complete medium and the autologous plasma prepared in step (1) for continued culture to obtain DC;
• (4) centrifuging the DC prepared in step (3) under conditions of 1500 rpm for 5 min, re-suspending and placing same in a serum-free XVIVO-15 medium comprising 1 mg/mL Perminede after completing the centrifugation, using Ad-P53 virus to transfect, and continuing to culture;
• and
• (5) re-suspending the DC cultured in step (4) in the normal saline to obtain a DC vaccine.