Comparison of the use of fresh PBMC versus fresh heparinised whole bloodpreclinical study dataset
We have data available from a comparison of the use of fresh peripheral blood mononuclear cells (PBMC) versus fresh heparinised whole blood.
Only when peptide pools as an antigen source are used PBMC exhibited a slightly higher stimulation capacity compared to whole blood. However, slightly higher background was observed when using PBMC to stimulate with antigen. Lowest background was observed when either peptides or protein were used for stimulation enabling assessment also of rare antigen-specific Th-cells below frequencies of 0.05% of peripheral CD4+ Th-cells. (Meier, Stark et al., 2008). We also tested usage of peptide pool preparations versus recombinant antigen for their stimulation capacity. While working with whole blood no differences in the capacity to elicit antigen-specific activation of Th-cells were observed, stimulation in PBMC induced higher activation (Meier, Stark et al., 2008).
We also evaluated the influence of the cell density on the stimulation outcome when PBMC were used for induction of Th-cell activation. Optimal Th-cell activation was induced when cultures were initiated with about 5x10E6/ml PBMC. Wrong adjustment of cell densities (e.g. to 1x10E6/ml reduced Th-cell activation to about 1/3 (Meier, Stark et al., 2008).