title: Serum-free process development: improving the yield and consistency of human mesenchymal stromal cell production.
abstract: OBJECTIVE
  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.affiliation: Centre for Biological Engineering, Loughborough University, Leicestershire, United Kingdom.
authors.forename: Thomas R J
authors.initials: TR
authors.lastname: Heathman
authors.affiliation: Centre for Biological Engineering, Loughborough University, Leicestershire, United Kingdom.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Stolzing
authors.affiliation: Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany; Translational Centre for Regenerative Medicine, Leipzig University, Leipzig, Germany.
authors.forename: Claire
authors.initials: C
authors.lastname: Fabian
authors.affiliation: 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.
authors.forename: Qasim A
authors.initials: QA
authors.lastname: Rafiq
authors.affiliation: Centre for Biological Engineering, Loughborough University, Leicestershire, United Kingdom.
authors.forename: Karen
authors.initials: K
authors.lastname: Coopman
authors.affiliation: Centre for Biological Engineering, Loughborough University, Leicestershire, United Kingdom; Centre for Bioprocess Engineering, University of Birmingham, Birmingham, United Kingdom.
authors.forename: Alvin W
authors.initials: AW
authors.lastname: Nienow
authors.affiliation: FUJIFILM Diosynth Biotechnologies, Billingham, United Kingdom.
authors.forename: Bo
authors.initials: B
authors.lastname: Kara
authors.affiliation: 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.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Hewitt
mesh_terms: D001854:Bone Marrow Cells / Q000166:cytology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D049109:Cell Proliferation; D048429:Cell Size; D003470:Culture Media / Q000737:chemistry; D015870:Gene Expression; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000502:physiology; D010012:Osteogenesis
chemical_list: D003470:Culture Media
keywords: cell-based therapy; comparability; consistency; human mesenchymal stromal cell; manufacturing; regenerative medicine; serum-free; yield