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

• Published in: Journal of tissue engineering and regenerative medicine Vol. 11; no. 5; pp. 1630 - 1640
• Main Authors: Soure, António M., Fernandes‐Platzgummer, Ana, Moreira, Francisco, Lilaia, Carla, Liu, Shi‐Hwei, Ku, Chen‐Peng, Huang, Yi‐Feng, Milligan, William, Cabral, Joaquim M. S., Silva, Cláudia L.
• Format: Journal Article
• Language: English
• Published: England Hindawi Limited 01.05.2017
• Subjects: Beads; Blood Platelets - chemistry; Cell adhesion; Cell adhesion & migration; Cell culture; Cell Separation - methods; Differentiation; ex vivo expansion; Flasks; Harvesting; human platelet lysate; Humans; Manufacturing; mesenchymal stem/stromal cells; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - metabolism; Mesenchyme; microcarriers; Plastics; Regenerative medicine; scale‐up; stem cell isolation; Stromal cells; Tissue engineering; Umbilical cord; Umbilical Cord - cytology; Umbilical Cord - metabolism; umbilical cord matrix; xenogeneic‐free; microcarriers; scale-up; mesenchymal stem/stromal cells; umbilical cord matrix; ex vivo expansion; human platelet lysate; xenogeneic-free; stem cell isolation
• ISSN: 1932-6254; 1932-7005
• DOI: 10.1002/term.2200

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.