title: Systematic microcarrier screening and agitated culture conditions improves human mesenchymal stem cell yield in bioreactors.
abstract: Production of human mesenchymal stem cells for allogeneic cell therapies requires scalable, cost-effective manufacturing processes. Microcarriers enable the culture of anchorage-dependent cells in stirred-tank bioreactors. However, no robust, transferable methodology for microcarrier selection exists, with studies providing little or no reason explaining why a microcarrier was employed. We systematically evaluated 13 microcarriers for human bone marrow-derived MSC (hBM-MSCs) expansion from three donors to establish a reproducible and transferable methodology for microcarrier selection. Monolayer studies demonstrated input cell line variability with respect to growth kinetics and metabolite flux. HBM-MSC1 underwent more cumulative population doublings over three passages in comparison to hBM-MSC2 and hBM-MSC3. In 100 mL spinner flasks, agitated conditions were significantly better than static conditions, irrespective of donor, and relative microcarrier performance was identical where the same microcarriers outperformed others with respect to growth kinetics and metabolite flux. Relative growth kinetics between donor cells on the microcarriers were the same as the monolayer study. Plastic microcarriers were selected as the optimal microcarrier for hBM-MSC expansion. HBM-MSCs were successfully harvested and characterised, demonstrating hBM-MSC immunophenotype and differentiation capacity. This approach provides a systematic method for microcarrier selection, and the findings identify potentially significant bioprocessing implications for microcarrier-based allogeneic cell therapy manufacture.
authors.affiliation: Centre for Biological Engineering, Department of Chemical Engineering, Loughborough University, Leicestershire, United Kingdom.|Wolfson School of Manufacturing and Mechanical 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, Department of Chemical Engineering, Loughborough University, Leicestershire, United Kingdom.
authors.forename: Karen
authors.initials: K
authors.lastname: Coopman
authors.affiliation: Centre for Biological Engineering, Department of Chemical Engineering, Loughborough University, Leicestershire, United Kingdom.|School of Chemical Engineering, University of Birmingham, Edgbaston, Birmingham, United Kingdom.
authors.forename: Alvin W
authors.initials: AW
authors.lastname: Nienow
authors.affiliation: Centre for Biological Engineering, Department of Chemical Engineering, Loughborough University, Leicestershire, United Kingdom. c.j.hewitt@aston.ac.uk.|Aston Medical Research Institute, School of Life and Health Sciences, Aston University, Aston Triangle, Birmingham, United Kingdom. c.j.hewitt@aston.ac.uk.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Hewitt
mesh_terms: D019149:Bioreactors; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*
keywords: Bioreactor; Cell therapy bioprocessing; Human mesenchymal stem cell; Microcarrier; Regenerative medicine