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An ultra scale-down analysis of the recovery by dead-end centrifugation of human cells for therapy.

Delahaye, M; Lawrence, K; Ward, SJ; Hoare, M; (2015) An ultra scale-down analysis of the recovery by dead-end centrifugation of human cells for therapy. Biotechnol Bioeng , 112 (5) 997 - 1011. 10.1002/bit.25519. Green open access

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Abstract

An ultra scale-down method is described to determine the response of cells to recovery by dead-end (batch) centrifugation under commercially defined manufacturing conditions. The key variables studied are the cell suspension hold time prior to centrifugation, the relative centrifugal force (RCF), time of centrifugation, cell pellet resuspension velocities, and number of resuspension passes. The cell critical quality attributes studied are the cell membrane integrity and the presence of selected surface markers. Greater hold times and higher RCF values for longer spin times all led to the increased loss of cell membrane integrity. However, this loss was found to occur during intense cell resuspension rather than the preceding centrifugation stage. Controlled resuspension at low stress conditions below a possible critical stress point led to essentially complete cell recovery even at conditions of extreme centrifugation (e.g., RCF of 10000 g for 30 mins) and long (~2 h) holding times before centrifugation. The susceptibility to cell loss during resuspension under conditions of high stress depended on cell type and the age of cells before centrifugation and the level of matrix crosslinking within the cell pellet as determined by the presence of detachment enzymes or possibly the nature of the resuspension medium. Changes in cell surface markers were significant in some cases but to a lower extent than loss of cell membrane integrity. Biotechnol. Bioeng. 2015;112: 997-1011. © 2014 Wiley Periodicals, Inc.

Type: Article
Title: An ultra scale-down analysis of the recovery by dead-end centrifugation of human cells for therapy.
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.1002/bit.25519
Publisher version: http://dx.doi.org/10.1002/bit.25519
Language: English
Additional information: © 2014 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals, Inc. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Keywords: cell quality, cell therapy, centrifugation, human cells, ultra scale-down
UCL classification: UCL
UCL > Provost and Vice Provost Offices > UCL BEAMS
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science
UCL > Provost and Vice Provost Offices > UCL BEAMS > Faculty of Engineering Science > Dept of Biochemical Engineering
URI: https://discovery.ucl.ac.uk/id/eprint/1467808
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