Journal of Clinical & Experimental Pharmacology

Standardized platelet releasate products for clinical applications in cell therapy: A mathematical approach and an innovative quality control test

Co-organized Event International Conference on Toxicology and Clinical Pharmacology & 2^nd International Conference on Generic Drugs and Biosimilars

December 14-16, 2017 Rome, Italy

Francesco Agostini, Cristina Durante, Alessandro Da Ponte, Monica Battiston, Elisabetta Lombardi, Giuseppe Astori, Davide Corpillo, Luca Biondi and Mario Mazzucato

CRO Aviano National Cancer Center, Italy
Vicenza Hospital, Italy
GEMFORLAB �?? ABLE Biosciences, Italy
Bracco Imaging, Italy

Scientific Tracks Abstracts: Clin Exp Pharmacol

Abstract:

Background: Expanded mesenchymal stem cells (MSC) can be utilized for advanced cell therapy. Growth factors can be extracted from human platelet concentrates by physical or chemical methods. Standardized animal-free ancillary materials are required to obtain expanded MSC (Good Manufacturing Practice). Pooling single-donor products reduces variability, but the minimal pool size was never determined. A method predicting biological activity of additives is presently lacking: NMR spectroscopy and MALDI-TOF mass spectrometry provide widespread molecular characterization of biological samples. Methods: Platelet-apheresis products were frozen and thawed to obtain platelet lysates (PL) or added with CaCl2 to produce Supernatant-rich-in-growth-factors (SRGF). Growth rates of MSC cultured in media containing PL or SRGF were compared. Concentrations of n=10 growth factors were measured by ELISA in n=44 single-donor SRGF. Data matrix was analyzed by a novel algorithm simulating pools (n=500) of single-donor data with growing sample size (from n=2 to 20) and estimating coefficient of variation (CV). For validation we measured a) the CV of growth factor concentrations in n=10 pools manufactured according to algorithm results, b) growth rates of MSC expanded by separate SRGF batches. NMR and MALDI-TOF spectra composition of single-donor PL and SRGF were analyzed. Results: SRGF promoted higher proliferation rate vs PL. Growth factor concentrations in single-donor SRGF showed high variability. In silico analysis suggested that pooling n=16 single-donor SRGF reduced CV below 20%: results were confirmed assessing CV of concentrations in real pools of n=16 single SRGF. Separate SRGF pools failed to differently affect MSC growth rate. NMR and MALDI-TOF spectroscopy demonstrated segregation between PL and SRGF products. Discussion: Results suggest that SRGF performs better than PL to stimulate MSC duplication. Our validated algorithm demonstrated that pooling n=16 single-donor SRGF products ameliorates consistency of biological activity of SRGF batches. NMR and MALDI-TOF could predict quality of media additives for cell therapy products. Recent Publications 1. Agostini F, Polesel J, Battiston M Lombardi E, Zanolin S et al. (2017) Standardization of platelet releasate products for clinical applications in cell therapy: a mathematical approach. J. Transl Med. 15(1):107. Doi: 10.1186/s12967-0171210-z. 2. Bernardi M, Agostini F, Chieregato K, Amati E, Durante C (2017) The production method affects the efficacy of platelet derivatives to expand mesenchymal stromal cells in vitro. J Transl 15(1):90. Doi: 10.1186/s12967-017-1185-9. 3. Borghese C, Agostini F, Durante C, Colombatti A, Mazzucato M, Aldinucci D (2016) Clinical-grade quality plateletrich plasma releasate (PRP-R/SRGF) from CaCl2 -activated platelet concentrates promoted expansion of mesenchymal stromal cells. Vox Sang. 111(2):197-205. Doi: 10.1111/vox.12405.

Biography :

Francesco Agostini is a Senior Scientist (Biologist) at the Aviano National Cancer Center (Aviano; PN, Italy). He studied at the University of Trieste where he obtained his PhD in Molecular Biomedicine in 2010. He is involved in Translational Research focusing his interest on GMP-compliant methods to expand mesenchymal stem cell for targeted drug delivery in the oncology field.