title: A novel filtration system for point of care washing of cellular therapy products.
abstract: The cell therapy industry would greatly benefit from a simple point of care solution to remove dimethylsulphoxide (DMSO) from small-volume thawed cell suspensions before injection. A novel dead-end filtration device has been designed and validated, which takes advantage of the higher density of thawed cell suspensions to remove the DMSO and protein impurities from the cell suspension without fouling the filter membrane. The filter was designed to avoid fluid circuits and minimize the surface area that is contacted by the cell suspension, thus reducing cell losses by design. The filtration process was established through optimization of the fluid flow configuration, backflush cycles and filter geometry. Overall, this novel filtration device allows for a 1 ml of thawed cryopreserved cell suspensions, containing 107cells of a fetal lung fibroblast cell line (MRC-5), to be washed in less than 30 min. More than 95% of the DMSO and up to 94% of the albumin-fluorescein-isothiocyanate content can be removed while the viable cell recovery is higher than 80%. It is also demonstrated that this system can be used for bone marrow-derived human mesenchymal stem cells with more than 73% cell recovery and 85% DMSO reduction. This is the first time that a dead end (normal) filtration process has been used to successfully wash high-density human cell suspensions. In practice, this novel solid-liquid separation technology fills the need for small-volume washing in closed processing systems for cellular therapies. Copyright © 2016 John Wiley & Sons, Ltd.
authors.affiliation: The Advanced Centre for Biochemical Engineering, Department of Biochemical Engineering, University College London, Bernard Katz Building, London, UK.
authors.forename: Rui
authors.initials: R
authors.lastname: Tostoes
authors.affiliation: Dodgson Associates Ltd, London, UK.
authors.forename: John R
authors.initials: JR
authors.lastname: Dodgson
authors.affiliation: The Advanced Centre for Biochemical Engineering, Department of Biochemical Engineering, University College London, Bernard Katz Building, London, UK.
authors.forename: Ben
authors.initials: B
authors.lastname: Weil
authors.affiliation: The Advanced Centre for Biochemical Engineering, Department of Biochemical Engineering, University College London, Bernard Katz Building, London, UK.
authors.forename: Spyridon
authors.initials: S
authors.lastname: Gerontas
authors.affiliation: The Advanced Centre for Biochemical Engineering, Department of Biochemical Engineering, University College London, Bernard Katz Building, London, UK.
authors.forename: Chris
authors.initials: C
authors.lastname: Mason
authors.affiliation: The Advanced Centre for Biochemical Engineering, Department of Biochemical Engineering, University College London, Bernard Katz Building, London, UK.
authors.forename: Farlan
authors.initials: F
authors.lastname: Veraitch
mesh_terms: D002460:Cell Line; D064987:Cell- and Tissue-Based Therapy / Q000295:instrumentation / Q000379:methods*; D015925:Cryopreservation / Q000379:methods; D003451:Cryoprotective Agents / Q000737:chemistry*; D004121:Dimethyl Sulfoxide / Q000737:chemistry*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D005374:Filtration / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D019095:Point-of-Care Systems
chemical_list: D003451:Cryoprotective Agents; D004121:Dimethyl Sulfoxide
keywords: cell delivery; cell manufacturing; cell processing; cell therapy; cell washing; filtration; point of care; stem cell processing