title: Integrated culture platform based on a human platelet lysate supplement for the isolation and scalable manufacturing of umbilical cord matrix-derived mesenchymal stem/stromal cells.
abstract: Umbilical cord matrix (UCM)-derived mesenchymal stem/stromal cells (MSCs) are promising therapeutic candidates for regenerative medicine settings. UCM MSCs have advantages over adult cells as these can be obtained through a non-invasive harvesting procedure and display a higher proliferative capacity. However, the high cell doses required in the clinical setting make large-scale manufacturing of UCM MSCs mandatory. A commercially available human platelet lysate-based culture supplement (UltraGROTM, AventaCell BioMedical) (5%(v/v)) was tested to effectively isolate UCM MSCs and to expand these cells under (1) static conditions, using planar culture systems and (2) stirred culture using plastic microcarriers in a spinner flask. The MSC-like cells were isolated from UCM explant cultures after 11 ± 2 days. After five passages in static culture, UCM MSCs retained their immunophenotype and multilineage differentiation potential. The UCM MSCs cultured under static conditions using UltraGROTM-supplemented medium expanded more rapidly compared with UCM MSCs expanded using a previously established protocol. Importantly, UCM MSCs were successfully expanded under dynamic conditions on plastic microcarriers using UltraGROTM-supplemented medium in spinner flasks. Upon an initial 54% cell adhesion to the beads, UCM MSCs expanded by >13-fold after 5-6 days, maintaining their immunophenotype and multilineage differentiation ability. The present paper reports the establishment of an easily scalable integrated culture platform based on a human platelet lysate supplement for the effective isolation and expansion of UCM MSCs in a xenogeneic-free microcarrier-based system. This platform represents an important advance in obtaining safer and clinically meaningful MSC numbers for clinical translation. Copyright © 2016 John Wiley & Sons, Ltd.
authors.affiliation: Department of Bioengineering and iBB-, Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal.
authors.forename: António M
authors.initials: AM
authors.lastname: de Soure
authors.affiliation: Department of Bioengineering and iBB-, Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal.
authors.forename: Ana
authors.initials: A
authors.lastname: Fernandes-Platzgummer
authors.affiliation: Department of Bioengineering and iBB-, Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal.
authors.forename: Francisco
authors.initials: F
authors.lastname: Moreira
authors.affiliation: Hospital São Francisco Xavier, Centro Hospitalar Lisboa Ocidental, Lisboa, Portugal.
authors.forename: Carla
authors.initials: C
authors.lastname: Lilaia
authors.affiliation: R&D, AventaCell BioMedical Co., Ltd., New Taipei City, Taiwan.
authors.forename: Shi-Hwei
authors.initials: SH
authors.lastname: Liu
authors.affiliation: R&D, AventaCell BioMedical Co., Ltd., New Taipei City, Taiwan.|R&D, AventaCell BioMedical Co., Ltd, Atlanta, GA, USA.
authors.forename: Chen-Peng
authors.initials: CP
authors.lastname: Ku
authors.affiliation: R&D, AventaCell BioMedical Co., Ltd., New Taipei City, Taiwan.|R&D, AventaCell BioMedical Co., Ltd, Atlanta, GA, USA.
authors.forename: Yi-Feng
authors.initials: YF
authors.lastname: Huang
authors.affiliation: R&D, AventaCell BioMedical Co., Ltd., New Taipei City, Taiwan.|R&D, AventaCell BioMedical Co., Ltd, Atlanta, GA, USA.
authors.forename: William
authors.initials: W
authors.lastname: Milligan
authors.affiliation: Department of Bioengineering and iBB-, Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal.
authors.forename: Joaquim M S
authors.initials: JMS
authors.lastname: Cabral
authors.affiliation: Department of Bioengineering and iBB-, Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal.
authors.forename: Cláudia L
authors.initials: CL
authors.lastname: da Silva
mesh_terms: D001792:Blood Platelets / Q000737:chemistry*; D002469:Cell Separation / Q000379:methods*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D014470:Umbilical Cord / Q000166:cytology* / Q000378:metabolism
keywords: ex vivo expansion; human platelet lysate; mesenchymal stem/stromal cells; microcarriers; scale-up; stem cell isolation; umbilical cord matrix; xenogeneic-free