Thomas R J Heathman1, Alexandra Stolzing1, Claire Fabian2, Qasim A Rafiq3, Karen Coopman1, Alvin W Nienow4, Bo Kara5, Christopher J Hewitt6. 1. Centre for Biological Engineering, Loughborough University, Leicestershire, United Kingdom. 2. Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany; Translational Centre for Regenerative Medicine, Leipzig University, Leipzig, Germany. 3. Centre for Biological Engineering, Loughborough University, Leicestershire, United Kingdom; Aston Medical Research Institute, School of Life and Health Sciences, Aston University, Aston Triangle, Birmingham, United Kingdom. 4. Centre for Biological Engineering, Loughborough University, Leicestershire, United Kingdom; Centre for Bioprocess Engineering, University of Birmingham, Birmingham, United Kingdom. 5. FUJIFILM Diosynth Biotechnologies, Billingham, United Kingdom. 6. Centre for Biological Engineering, Loughborough University, Leicestershire, United Kingdom; Aston Medical Research Institute, School of Life and Health Sciences, Aston University, Aston Triangle, Birmingham, United Kingdom. Electronic address: c.j.hewitt@aston.ac.uk.
Abstract
BACKGROUND AIMS: The cost-effective production of human mesenchymal stromal cells (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 (BM-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 through a correlation (R(2) = 0.8705) across all conditions. PRIME-XV SFM has also shown increased consistency in BM-hMSC characteristics such as per cell metabolite utilization, 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.
BACKGROUND AIMS: The cost-effective production of human mesenchymal stromal cells (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 (BM-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 through a correlation (R(2) = 0.8705) across all conditions. PRIME-XV SFM has also shown increased consistency in BM-hMSC characteristics such as per cell metabolite utilization, 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.
Authors: Qasim A Rafiq; Mariana P Hanga; Thomas R J Heathman; Karen Coopman; Alvin W Nienow; David J Williams; Christopher J Hewitt Journal: Biotechnol Bioeng Date: 2017-07-27 Impact factor: 4.530
Authors: Ali Nassif; Benjamin Philippe Fournier; Ihsène Taihi; Caroline Pilon; José Cohen; Ariane Berdal; Bruno Gogly Journal: Stem Cell Res Ther Date: 2022-03-25 Impact factor: 6.832