A portion of pulsed and unpulsed mature dendritic cells will be frozen and used to monitor immunological responses elicited by the DC vaccine and transferred T cells during the clinical trial.

Fc-block for 30 min on ice

On day 7 of culture 35/40 x 10e6 mature peptide pulsed DC are frozen at 5 x 10e6 cells per vial in freezing medium as above.

The majority of monocytes (1 x 10e9) and all lymphocytes (6,4 x 10e9) are frozen in 90% A-plasma and 10% GMP-grade DMSO.

Take supernatant and freeze at -80°C for analysis of DC-stimulation

After thawing cells showed good viability (>90%) and the recovery was approximately 80%.