Author:

Keywords:

bioreactor, mesenchymal stromal cell expansion, Science & Technology, Life Sciences & Biomedicine, Cell & Tissue Engineering, Biotechnology & Applied Microbiology, Cell Biology, Hematology, Medicine, Research & Experimental, Research & Experimental Medicine, osteogenesis, quality control, process reproducibility, process monitoring and control, MESENCHYMAL STROMAL CELLS, ECTOPIC BONE-FORMATION, STEM-CELLS, THERAPY, DIFFERENTIATION, FIBRONECTIN, SPECIFICITY, STABILITY, MODEL, Adult, Animals, Bioreactors, Bone and Bones, Cell Culture Techniques, Cell Differentiation, Cell Proliferation, Cell- and Tissue-Based Therapy, Female, Humans, Male, Mice, Nude, Middle Aged, Periosteum, Stem Cells, Young Adult, 1103 Clinical Sciences, Immunology, 3204 Immunology, 3206 Medical biotechnology

Abstract:

Background aims. With the increasing scale in stem cell production, a robust and controlled cell expansion process becomes essential for the clinical application of cell-based therapies.The objective of this work was the assessment of a hollow fiber bioreactor (Quantum Cell Expansion System from Terumo BCT) as a cell production unit for the clinical-scale production of human periosteum derived stem cells (hPDCs). Methods. We aimed to demonstrate comparability of bioreactor production to standard culture flask production based on a product characterization in line with the International Society of Cell Therapy in vitro benchmarks and supplemented with a compelling quantitative in vivo bone-forming potency assay. Multiple process read-outs were implemented to track process performance and deal with donor-to-donor-related variation in nutrient needs and harvest timing. Results. The data show that the hollow fiber bioreactor is capable of robustly expanding autologous hPDCs on a clinical scale (yield between 316 million and 444 million cells starting from 20 million after ± 8 days of culture) while maintaining their in vitro quality attributes compared with the standard flask-based culture. The in vivo boneforming assay on average resulted in 10.3 ± 3.7% and 11.0 ± 3.8% newly formed bone for the bioreactor and standard culture flask respectively.The analysis showed that the Quantum system provides a reproducible cell expansion process in terms of yields and culture conditions for multiple donors.