Sabrina Viau1, Anaïs Lagrange2, Lucie Chabrand2, Judith Lorant2, Marine Charrier3, Karl Rouger4, Ignacio Alvarez5, Sandy Eap2, Bruno Delorme2. 1. Biotherapy Division, Macopharma, Mouvaux, France. Electronic address: sabrina.viau@macopharma.com. 2. Biotherapy Division, Macopharma, Mouvaux, France. 3. PAnTher, Institut National de la Recherche Agronomique (INRA), Ecole Nationale Vétérinaire, Agro-alimentaire et de l'Alimentation Nantes-Atlantique (Oniris), Université Bretagne Loire, Nantes, France; Université de Nantes, Université Bretagne Loire, Nantes, France. 4. PAnTher, Institut National de la Recherche Agronomique (INRA), Ecole Nationale Vétérinaire, Agro-alimentaire et de l'Alimentation Nantes-Atlantique (Oniris), Université Bretagne Loire, Nantes, France. 5. Medical Department, Macopharma, Tourcoing, France.
Abstract
BACKGROUND: Human platelet lysate (hPL) represents a powerful alternative to fetal bovine serum (FBS) for human mesenchymal stromal cell (hMSC) expansion. However, the large variability in hPL sources and production protocols gives rise to discrepancies in product quality, characterization and poor batch-to-batch standardization. METHODS: hPL prepared with more than 200 donors (200+DhPL) or with five donors (5DhPL) were compared in terms of growth factor (GF) contents and biochemical analysis. A multiple protein assay and proteomic analysis were performed to further characterize 200+DhPL batches. We also compared the phenotypic and functional characteristics of bone marrow (BM)-hMSCs grown in 200+DhPL versus FBS+basic fibroblast growth factor (bFGF). RESULTS: By contrast to 5DhPL, industrial 200+DhPL displayed a strong standardization of GF contents and biochemical characteristics. We identified specific plasmatic components and platelet-released factors as the most relevant markers for the evaluation of the standardization of hPL batches. We used a multiplex assay and proteomic analysis of 200+DhPL to establish a proteomic signature and demonstrated the robust standardization of batches. 200+DhPL was shown to improve and standardize BM-hMSC expansion compared with FBS+bFGF. The levels of expression of BM-hMSC membrane markers were found to be much more homogeneous between batches when cells were cultured in 200+DhPL. BM-hMSCs cultured in parallel under both conditions displayed similar adipogenic and osteogenic differentiation potential and immunosuppressive properties. CONCLUSIONS: We report a standardization of hPL and the importance of such standardization for the efficient amplification of more homogeneous and reproducible cell therapy products.
BACKGROUND:Human platelet lysate (hPL) represents a powerful alternative to fetal bovine serum (FBS) for human mesenchymal stromal cell (hMSC) expansion. However, the large variability in hPL sources and production protocols gives rise to discrepancies in product quality, characterization and poor batch-to-batch standardization. METHODS:hPL prepared with more than 200 donors (200+DhPL) or with five donors (5DhPL) were compared in terms of growth factor (GF) contents and biochemical analysis. A multiple protein assay and proteomic analysis were performed to further characterize 200+DhPL batches. We also compared the phenotypic and functional characteristics of bone marrow (BM)-hMSCs grown in 200+DhPL versus FBS+basic fibroblast growth factor (bFGF). RESULTS: By contrast to 5DhPL, industrial 200+DhPL displayed a strong standardization of GF contents and biochemical characteristics. We identified specific plasmatic components and platelet-released factors as the most relevant markers for the evaluation of the standardization of hPL batches. We used a multiplex assay and proteomic analysis of 200+DhPL to establish a proteomic signature and demonstrated the robust standardization of batches. 200+DhPL was shown to improve and standardize BM-hMSC expansion compared with FBS+bFGF. The levels of expression of BM-hMSC membrane markers were found to be much more homogeneous between batches when cells were cultured in 200+DhPL. BM-hMSCs cultured in parallel under both conditions displayed similar adipogenic and osteogenic differentiation potential and immunosuppressive properties. CONCLUSIONS: We report a standardization of hPL and the importance of such standardization for the efficient amplification of more homogeneous and reproducible cell therapy products.
Authors: Siddharth Shanbhag; Samih Mohamed-Ahmed; Turid Helen Felli Lunde; Salwa Suliman; Anne Isine Bolstad; Tor Hervig; Kamal Mustafa Journal: Stem Cell Res Ther Date: 2020-09-23 Impact factor: 6.832