ArticleCancer immunology, immunotherapy : CII2013
Serum-free freezing media support high cell quality and excellent ELISPOT assay performance across a wide variety of different assay protocols.
Article in Cancer immunology, immunotherapy : CII, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed, 28 citations in OpenAlex.
- T cell immunomonitoring: a comparative analysis of traditional and novel methods to quantify and characterize human antigen-specific T cells.Nature protocols · 2026Review
- Development of a new cell isolation device FlowMagicTM.PloS one · 2025Article
- Technical pitfalls when collecting, cryopreserving, thawing, and stimulating human T-cells.Frontiers in immunology · 2024Review
- Optimization of Peripheral Blood Mononuclear Cell Processing for Improved Clinical ELISpot Assay Performance.The AAPS journal · 2023Article
- Monitoring cell-mediated immune responses in AAV gene therapy clinical trials using a validated IFN-γ ELISpot method.Molecular therapy. Methods & clinical development · 2021Article
- Article
- Optimization of a Method to Detect Autoantigen-Specific T-Cell Responses in Type 1 Diabetes.Frontiers in immunology · 2020Article
- The effects of storage temperature on PBMC gene expression.BMC immunology · 2016Article
- Toward harmonized phenotyping of human myeloid-derived suppressor cells by flow cytometry: results from an interim study.Cancer immunology, immunotherapy : CII · 2016Article
- Cryopreservation of Human Mucosal Leukocytes.PloS one · 2016Article
- Data analysis as a source of variability of the HLA-peptide multimer assay: from manual gating to automated recognition of cell clusters.Cancer immunology, immunotherapy : CII · 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
- 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
- CIMT 2014: Next waves in cancer immunotherapy--report on the 12th annual meeting of the Association for Cancer Immunotherapy: May 6–8 2014, Mainz, Germany.Human vaccines & immunotherapeutics · 2014Article
- Immunological Monitoring to Rationally Guide AAV Gene Therapy.Frontiers in immunology · 2013Review
- Vaccine against autoimmune disease: antigen-specific immunotherapy.Current opinion in immunology · 2013Review
Corrections and comments
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Authors and funding
11 authors at 5 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Robust and sensitive ELISPOT protocols are commonly applied concomitant with the development of new immunotherapeutics. Despite the knowledge that individual serum batches differ in their composition and may change properties over time, serum is still commonly used in immunologic assays. Commercially available serum batches are expensive, limited in quantity and need to be pretested for suitability in immunologic assays, which is a laborious process. The aim of this study was to test whether serum-free freezing media can lead to high cell viability and favorable performance across multiple ELISPOT assay protocols. Thirty-one laboratories from ten countries participated in a proficiency panel organized by the Cancer Immunotherapy Immunoguiding Program to test the influence of different freezing media on cell quality and immunologic function. Each center received peripheral blood mononuclear cells which were frozen in three different media. The participants were asked to quantify antigen-specific CD8+ T-cell responses against model antigens using their locally established IFN-gamma ELISPOT protocols. Self-made and commercially available serum-free freezing media led to higher cell viability and similar cell recovery after thawing and resting compared to freezing media supplemented with human serum. Furthermore, the test performance as determined by (1) background spot production, (2) replicate variation, (3) frequency of detected antigen-specific spots and (4) response detection rate was similar for serum and serum-free conditions. We conclude that defined and accessible serum-free freezing media should be recommended for freezing cells stored for subsequent ELISPOT analysis.
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