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

Table 1 DC viability following culture in four different media.

	Number of viable DCs harvested from 5 × 10⁶ monocytes per T25 cm2 flask
Media condition	Before lysate-loading and maturation (10⁶)	After lysate-loading and maturation (10⁶)	% viable DC generated (± SEM)
AIM-V no AB serum	4.52 ± 0.88	2.29 ± 0.36	53.39 ± 5.94
AIM-V + 2% AB serum	5.10 ± 0.92	3.05 ± 0.3	64.72 ± 7.14
CellGenix no AB serum	5.09 ± 0.61	2.76 ± 0.87	43.64 ± 6.17
CellGenix + 2% AB serum	5.48 ± 0.67	3.87 ± 0.35	73.28 ± 7.58*

CellGenix DC media supplemented with 2% human AB serum yielded the highest percentage of viable DCs after loading with HOCl-oxidized lysate (1 DC to 1 tumor cell ratio for 20-24 h) and 16 h maturation with LPS (60 EU/ml) and IFN-γ (2000 IU/ml). The final percentage of viable DC generated was calculated as follows: (number of viable DCs obtained per flask after lysate-loading and maturation ÷ number of viable DCs obtained per flask before lysate-loading and maturation) × 100%. The mean of 6 independent experiments ± standard error of the mean [SEM]) is shown. *P = 0.016; final % of viable DCs generated from CellGenix + 2% AB serum was significantly different from the final % of viable DCs generated with AIM-V no AB serum, AIM-V + 2% AB serum or CellGenix no AB serum (one-way ANOVA).

ISSN: 1479-5876