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Table 2 DC viability following two different harvesting methods.

From: Optimizing parameters for clinical-scale production of high IL-12 secreting dendritic cells pulsed with oxidized whole tumor cell lysate

Harvesting method

% viable DC (± SEM)

Trypsinization

79.94 ± 3.66 ***

Cold PBS + scraping

59.47 ± 2.44

  1. Trypsinization method yielded higher % of viable DCs compared to harvesting DCs with cold DPBS and scraping. Fresh monocytes were cultured in CellGenix DC media supplemented with 2% human AB serum for 4 days. Then immature DCs were loaded with oxidized tumor lysate for 20 to 24 h followed by maturation with LPS and IFN-γ for 16 h. The percentage of viable DC was determined by Trypan blue exclusion using the following formula: (total number of live non-Trypan blue DCs in all the four big squares of the Bürker chamber ÷ total number of live and dead DCs counted in the four big squares of the Bürker chamber) × 100%. Data is mean of 3 independent experiments (± SEM). *** P = 0.0003; highly significant difference was observed from cold DPBS + scraping (unpaired Student's t test).