Performance of serum-supplemented and serum-free media in IFNgamma Elispot Assays for human T cells.

Journal Article (Journal Article)

The choice of serum for supplementation of media for T cell assays and in particular, Elispot has been a major challenge for assay performance, standardization, optimization, and reproducibility. The Assay Working Group of the Cancer Vaccine Consortium (CVC-CRI) has recently identified the choice of serum to be the leading cause for variability and suboptimal performance in large international Elispot proficiency panels. Therefore, a serum task force was initiated to compare the performance of commercially available serum-free media to laboratories' own medium/serum combinations. The objective of this project was to investigate whether a serum-free medium exists that performs as well as lab-own serum/media combinations with regard to antigen-specific responses and background reactivity in Elispot. In this way, a straightforward solution could be provided to address the serum challenge. Eleven laboratories tested peripheral blood mononuclear cells (PBMC) from four donors for their reactivity against two peptide pools, following their own Standard Operating Procedure (SOP). Each laboratory performed five simultaneous experiments with the same SOP, the only difference between the experiments was the medium used. The five media were lab-own serum-supplemented medium, AIM-V, CTL, Optmizer, and X-Vivo. The serum task force results demonstrate compellingly that serum-free media perform as well as qualified medium/serum combinations, independent of the applied SOP. Recovery and viability of cells are largely unaffected by serum-free conditions even after overnight resting. Furthermore, one serum-free medium was identified that appears to enhance antigen-specific IFNgamma-secretion.

Full Text

Duke Authors

Cited Authors

  • Janetzki, S; Price, L; Britten, CM; van der Burg, SH; Caterini, J; Currier, JR; Ferrari, G; Gouttefangeas, C; Hayes, P; Kaempgen, E; Lennerz, V; Nihlmark, K; Souza, V; Hoos, A

Published Date

  • April 2010

Published In

Volume / Issue

  • 59 / 4

Start / End Page

  • 609 - 618

PubMed ID

  • 19894047

Pubmed Central ID

  • 19894047

Electronic International Standard Serial Number (EISSN)

  • 1432-0851

Digital Object Identifier (DOI)

  • 10.1007/s00262-009-0788-2


  • eng

Conference Location

  • Germany