Suspension culture on microcarriers and as aggregates enables expansion and differentiation of pluripotent stem cells (PSCs) |
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| Institution: | 1. Unité de thérapie Cellulaire et banque de tissus, Université de Lorraine, CHRU-Nancy, Nancy, France;2. Université de Lorraine, UMR CNRS UL 7365, Campus Santé, Nancy, France;3. Département Méthodologie, Promotion, Investigation, Université de Lorraine, CHRU-Nancy, Nancy, France;4. llogeneic Stem Cell Transplantation Department, Université de Lorraine, CHRU-Nancy, Nancy, France.;5. Flow Cytometry Platform, Batiment de Biologie, Université de Lorraine, CHRU-Nancy, Nancy, France;1. Faculty of Health Sciences, University of Ottawa, Ottawa, Ontario, Canada;2. Faculty of Medicine, University of Ottawa, Ottawa, Ontario, Canada;3. Faculty of Medicine, University of Toronto, Toronto, Ontario, Canada;4. Ottawa Hospital Research Institute, Ottawa, Ontario, Canada;5. Stem Cells, Canadian Blood Services, Ottawa, Ontario, Canada;6. Department of Medicine, The Ottawa Hospital, Ottawa, Ontario, Canada;1. Aenitis Technologies, Paris, France;2. Institut André Lwoff, INSERM UMR-MD 1197, Villejuif, France;3. Laboratoire PMMH, UMR7636 CNRS, ESPCI Paris - PSL, Paris Sciences Lettres, Sorbonne Université, Paris, France;4. Université de Paris, Assistance Publique-Hôpitaux de Paris, Hôpital Saint-Louis, Paris, France;5. Unité de Thérapie Cellulaire, INSERM U976, Centre d''investigation clinique de Biothérapies CBT501, Paris, France;6. Institut de Recherche Biomédicale des Armées, Clamart, France;7. Centre de Transfusion Sanguine des Armées, Clamart, France;1. Centre for Regenerative Medicine Research, School of Pharmacy and Bioengineering, Keele University, Keele, Newcastle, United Kingdom;2. Robert Jones and Agnes Hunt Orthopaedic Hospital, Gobowen, Oswestry, Shropshire, United Kingdom;1. Preclinical Safety Department, Translational Medicine, Novartis Institutes for BioMedical Research, Basel, Switzerland;2. Division of Molecular and Systems Toxicology, Department of Pharmaceutical Sciences, University of Basel, Basel, Switzerland;3. Chemical Biology and Therapeutics Department, Novartis Institutes for BioMedical Research, Basel, Switzerland |
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| Abstract: | Background aimsHuman pluripotent stem cells (PSCs) hold a great promise for promoting regenerative medical therapies due to their ability to generate multiple mature cell types and for their high expansion potential. However, cell therapies require large numbers of cells to achieve desired therapeutic effects, and traditional two-dimensional static culture methods cannot meet the required production demand for cellular therapies. One solution to this problem is scaling up expansion of PSCs in bioreactors using culture strategies such as growing cells on microcarriers or as aggregates in suspension culture.MethodsIn this study, we directly compared PSC expansion and quality parameters in microcarrier- and aggregate-cultures grown in single-use vertical-wheel bioreactors.ResultsWe showed comparable expansion of cells on microcarriers and as aggregates by day 6 with a cell density reaching 2.2 × 106 cells/mL and 1.8 × 106 cells/mL and a fold-expansion of 22- and 18-fold, respectively. PSCs cultured on microcarriers and as aggregates were comparable with parallel two-dimensional cultures and with each other in terms of pluripotency marker expression and retention of other pluripotency characteristics as well as differentiation potential into three germ layers, neural precursor cells and cardiomyocytes.ConclusionsOur study did not demonstrate a clear advantage between the two three-dimensional methods for the quality parameters assessed. This analysis adds support to the use of bioreactor systems for large scale expansion of PSCs, demonstrating that the cells retain key characteristics of PSCs and differentiation potential in suspension culture. |
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