Serum-free process development: improving the yield and consistency of human mesenchymal stem cell production Thomas Heathman Alexandra Stolzing Claire Fabian Qasim A. Rafiq Alvin W. Nienow Karen Coopman Bo Kara Christopher J. Hewitt 2134/18932 https://repository.lboro.ac.uk/articles/journal_contribution/Serum-free_process_development_improving_the_yield_and_consistency_of_human_mesenchymal_stem_cell_production/9563147 Background: The cost effective production of hMSCs for off-the-shelf and patient specific therapies will require an increasing focus on improving product yield and driving manufacturing consistency. Methods: Bone-marrow derived hMSCs from two donors were expanded for 36 days in monolayer with medium supplemented with either fetal bovine serum (FBS) or PRIME-XV® Serum-free Medium (SFM). Cells were assessed throughout culture for proliferation, mean cell diameter, colony forming potential, osteogenic potential, gene expression and metabolites. Results: Expansion of BM-hMSCs in PRIME-XV® SFM resulted in a significantly higher growth rate (p < 0.001) and increased consistency between donors compared with FBS-based culture. FBS-based culture showed an inter batch production range of 0.9 and 5 days per dose compared with 0.5 and 0.6 days in SFM for each BM-hMSC donor line. The consistency between donors was also improved by the use of PRIME-XV® SFM, with a production range of 0.9 days compared with 19.4 days in FBS-based culture. Mean cell diameter has also been demonstrated as a process metric for BM-hMSC growth rate and senescence via a correlation (R2 = 0.8705) across all conditions. PRIME-XV® SFM has also shown increased consistency in BM-hMSC characteristics such as per cell metabolite utilisation, in vitro colony forming potential and osteogenic potential despite the higher number of population doublings. Conclusions: We have increased the yield and consistency of BM-hMSC expansion between donors, demonstrating a level of control over the product, which has the potential to increase the cost effectiveness and reduce the risk in these manufacturing processes. 2015-10-02 11:45:29 Serum-free Consistency Human mesenchymal stem cell Manufacturing Cell-based therapy Regenerative medicine Yield Comparability Mechanical Engineering not elsewhere classified