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- Title
Validation of an ICH Q2 Compliant Flow Cytometry-Based Assay for the Assessment of the Inhibitory Potential of Mesenchymal Stromal Cells on T Cell Proliferation.
- Authors
Piede, Natascha; Bremm, Melanie; Farken, Anne; Pfeffermann, Lisa-Marie; Cappel, Claudia; Bonig, Halvard; Fingerhut, Theres; Puth, Laura; Vogelsang, Kathrin; Peinelt, Andreas; Marschalek, Rolf; Müller, Matthias; Bader, Peter; Kuçi, Zyrafete; Kuçi, Selim; Huenecke, Sabine
- Abstract
Mesenchymal stromal cells (MSCs) have the potential to suppress pathological activation of immune cells and have therefore been considered for the treatment of Graft-versus-Host-Disease. The clinical application of MSCs requires a process validation to ensure consistent quality. A flow cytometry-based mixed lymphocyte reaction (MLR) was developed to analyse the inhibitory effect of MSCs on T cell proliferation. Monoclonal antibodies were used to stimulate T cell expansion and determine the effect of MSCs after four days of co-culture based on proliferation tracking with the violet proliferation dye VPD450. Following the guidelines of the International Council for Harmonisation (ICH) Q2 (R1), the performance of n = 30 peripheral blood mononuclear cell (PBMC) donor pairs was assessed. The specific inhibition of T cells by viable MSCs was determined and precision values of <10% variation for repeatability and <15% for intermediate precision were found. Compared to a non-compendial reference method, a linear correlation of r = 0.9021 was shown. Serial dilution experiments demonstrated a linear range for PBMC:MSC ratios from 1:1 to 1:0.01. The assay was unaffected by PBMC inter-donor variability. In conclusion, the presented MLR can be used as part of quality control tests for the validation of MSCs as a clinical product.
- Subjects
MONONUCLEAR leukocytes; T cells; STROMAL cells; CELL proliferation
- Publication
Cells (2073-4409), 2023, Vol 12, Issue 6, p850
- ISSN
2073-4409
- Publication type
Article
- DOI
10.3390/cells12060850