Using the definition above 9/10 PBMC samples were responsive to the CMV and 9/10 to the CEF peptide pool (Table 2), independent of the storage condition. Also, with 0–12 spot-forming cells per 106 PBMC, the background was very low, independent of the sample storage (data not shown). The results indicated that repeated temperature fluctuation during sample storage decreases the antigen-specific immune response of T-cells measured by IFN-γ ELISpot (Fig. 7). We detected only a small decrease in T-cell functionality using the protective hood system. Using this system we detected a mean reduction of −6.54% (±15.89) in response

to CEF peptide pool antigen-stimulation, ranging from +5.60% to −37.85% for different donors. A similar average decrease of −4.36% (±8.24) in T-cell function after T-cell stimulation using the CMV peptide pool was also detectable. The differences in FK506 mw CMV specific immune responses ranged from +6.25% to −15.12%. In contrast, a strong reduction in the immune response was detected for samples

exposed to temperature fluctuations, with cyclical temperature Selleckchem Dabrafenib rises towards room temperature, when compared to samples stored without any temperature cycling. Repeated sample storage and removal without the use of the protected hood system led to an average decrease of −29.71% (±25.36) in response to the CEF peptide pool and −28.02% (±20.69) after antigene stimulation with the CMV peptide pool as. In comparison,

in samples stored without temperature fluctuation the reduction ranged from +3.09% to −44.38% and from −0.89% to −66.24% in response to the CEF and CMV peptide pool, respectively. In summary, these results show that the maintenance of cell viability, recovery and T-cell functionality is strongly dependent on maintaining the samples in storage conditions without temperature fluctuations. Repeated temperature shifts led to a decrease in all measured parameters. Cryopreservation of cells offers many advantages to the research community, such as banking of multiple aliquots of cells from multicenter studies of large cohorts of individuals. It allows precious samples to be available for future studies, often using newly developed techniques or assays. 4-Aminobutyrate aminotransferase Additionally, samples of the same donor banked over time can be simultaneously processed, allowing greater inter- and intra-laboratory control and reducing costs. High-quality and reproducible cryopreservation of specimens is extremely important and demanding for the success of these studies. Cryopreservation can have significant effects and on PBMC viability, recovery and functionality [39] and [49] and many parameters are known to influence recovery including population purity, processing time, freezing medium, thawing and overnight culture conditions [5], [9], [12], [14], [21], [24] and [36].