title: Scalability and process transfer of mesenchymal stromal cell production from monolayer to microcarrier culture using human platelet lysate.
abstract: OBJECTIVE
  The selection of medium and associated reagents for human mesenchymal stromal cell (hMSC) culture forms an integral part of manufacturing process development and must be suitable for multiple process scales and expansion technologies.
  
  
  METHODS
  In this work, we have expanded BM-hMSCs in fetal bovine serum (FBS)- and human platelet lysate (HPL)-containing media in both a monolayer and a suspension-based microcarrier process.
  
  
  RESULTS
  The introduction of HPL into the monolayer process increased the BM-hMSC growth rate at the first experimental passage by 0.049 day and 0.127/day for the two BM-hMSC donors compared with the FBS-based monolayer process. This increase in growth rate in HPL-containing medium was associated with an increase in the inter-donor consistency, with an inter-donor range of 0.406 cumulative population doublings after 18 days compared with 2.013 in FBS-containing medium. Identity and quality characteristics of the BM-hMSCs are also comparable between conditions in terms of colony-forming potential, osteogenic potential and expression of key genes during monolayer and post-harvest from microcarrier expansion. BM-hMSCs cultured on microcarriers in HPL-containing medium demonstrated a reduction in the initial lag phase for both BM-hMSC donors and an increased BM-hMSC yield after 6 days of culture to 1.20 ± 0.17 × 10(5) and 1.02 ± 0.005 × 10(5) cells/mL compared with 0.79 ± 0.05 × 10(5) and 0.36 ± 0.04 × 10(5) cells/mL in FBS-containing medium.
  
  
  CONCLUSIONS
  This study has demonstrated that HPL, compared with FBS-containing medium, delivers increased growth and comparability across two BM-hMSC donors between monolayer and microcarrier culture, which will have key implications for process transfer during scale-up.
authors.affiliation: PCT, a Caladrius Company, NJ, USA; Centre for Biological Engineering, Loughborough University, Leicestershire, UK.
authors.forename: Thomas R J
authors.initials: TR
authors.lastname: Heathman
authors.affiliation: Centre for Biological Engineering, Loughborough University, Leicestershire, UK; Interdisciplinary Centre for Bioinformatics, University of Leipzig, Leipzig, Germany.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Stolzing
authors.affiliation: Interdisciplinary Centre for Bioinformatics, University of Leipzig, Leipzig, Germany; Fraunhofer Institute for Cell Therapy and Immunology, Leipzig, Germany.
authors.forename: Claire
authors.initials: C
authors.lastname: Fabian
authors.affiliation: Centre for Biological Engineering, Loughborough University, Leicestershire, UK; Aston Medical Research Institute, School of Life and Health Sciences, Aston University, Aston Triangle, Birmingham, UK.
authors.forename: Qasim A
authors.initials: QA
authors.lastname: Rafiq
authors.affiliation: Centre for Biological Engineering, Loughborough University, Leicestershire, UK.
authors.forename: Karen
authors.initials: K
authors.lastname: Coopman
authors.affiliation: Centre for Biological Engineering, Loughborough University, Leicestershire, UK; Aston Medical Research Institute, School of Life and Health Sciences, Aston University, Aston Triangle, Birmingham, UK; Centre for Bioprocess Engineering, University of Birmingham, UK.
authors.forename: Alvin W
authors.initials: AW
authors.lastname: Nienow
authors.affiliation: FUJIFILM Diosynth Biotechnologies, Billingham, UK.
authors.forename: Bo
authors.initials: B
authors.lastname: Kara
authors.affiliation: Centre for Biological Engineering, Loughborough University, Leicestershire, UK; Aston Medical Research Institute, School of Life and Health Sciences, Aston University, Aston Triangle, Birmingham, UK. Electronic address: c.j.hewitt@aston.ac.uk.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Hewitt
mesh_terms: D000818:Animals; D001792:Blood Platelets / Q000166:cytology*; D002417:Cattle; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002478:Cells, Cultured; D006801:Humans; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D055616:Microtechnology / Q000379:methods*; D054457:Tissue Scaffolds; D057170:Translational Research, Biomedical
keywords: bioprocess; cell-based therapy; comparability; harvest; human platelet lysate; manufacture; mesenchymal stromal cell; microcarrier expansion; process development; process transfer