ArticleCancer immunology, immunotherapy : CII2010
Performance of serum-supplemented and serum-free media in IFNgamma Elispot Assays for human T cells.
Article in Cancer immunology, immunotherapy : CII, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers.
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Who cites it
29 citing papers in PubMed.
- Trial
- T cell immunomonitoring: a comparative analysis of traditional and novel methods to quantify and characterize human antigen-specific T cells.Nature protocols · 2026Review
- A systematic approach towards long-term, serum-free cultivation of fish cells with the RTgill-W1 cell line as example.NAM journal · 2025Article
- Performance and Stability of New Class of Fetal Bovine Sera (FBS) and Its Lyophilized Form in ELISpot and FluoroSpot Assays: Applications for Monitoring the Immune Response in Vaccine, and Cell and Gene Immunotherapy in Clinical Trials.Methods in molecular biology (Clifton, N.J.) · 2024Article
- Optimization of Peripheral Blood Mononuclear Cell Processing for Improved Clinical ELISpot Assay Performance.The AAPS journal · 2023Article
- Evaluation of Cellular Immune Response to Adeno-Associated Virus-Based Gene Therapy.The AAPS journal · 2023Review
- Validation of the performance and suitability of a new class of FBS optimized for use in single-cell functional assays.Journal of immunological methods · 2023Review
- Monitoring cell-mediated immune responses in AAV gene therapy clinical trials using a validated IFN-γ ELISpot method.Molecular therapy. Methods & clinical development · 2021Article
- Influence of various medium environment to in vitro human T cell culture.In vitro cellular & developmental biology. Animal · 2018Article
- Extracellular milieu grossly alters pathogen-specific immune response of mammary epithelial cells.BMC veterinary research · 2015Article
- Guidelines for the automated evaluation of Elispot assays.Nature protocols · 2015Article
- Self-adjuvanted mRNA vaccination in advanced prostate cancer patients: a first-in-man phase I/IIa study.Journal for immunotherapy of cancer · 2015Article
- Article
- Article
- Harmonisation of short-term in vitro culture for the expansion of antigen-specific CD8(+) T cells with detection by ELISPOT and HLA-multimer staining.Cancer immunology, immunotherapy : CII · 2014Article
- Article
- Enzyme-linked immunospot assay for detection of human respiratory syncytial virus f protein-specific gamma interferon-producing T cells.Clinical and vaccine immunology : CVI · 2014Article
- Characterization of T-cell responses to cryptic epitopes in recipients of a noncodon-optimized HIV-1 vaccine.Journal of acquired immune deficiency syndromes (1999) · 2014Article
- Immunological Monitoring to Rationally Guide AAV Gene Therapy.Frontiers in immunology · 2013Review
- A harmonized approach to intracellular cytokine staining gating: Results from an international multiconsortia proficiency panel conducted by the Cancer Immunotherapy Consortium (CIC/CRI).Cytometry. Part A : the journal of the International Society for Analytical Cytology · 2013Article
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Authors and funding
14 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
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.
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