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A Quality-by-Design approach to risk reduction and optimization for human embryonic stem cell cryopreservation processes
journal contribution
posted on 2017-08-21, 08:37 authored by Peter Mitchell, Elizabeth RatcliffeElizabeth Ratcliffe, Paul Hourd, David Williams, Rob ThomasRob ThomasIt is well documented that cryopreservation and resuscitation of human embryonic stem cells (hESCs) is complex and ill-defined, and often suffers poor cell recovery and increased levels of undesirable cell differentiation. In this study we have applied Quality-by-Design (QbD) concepts to the critical processes of slow-freeze cryopreservation and resuscitation of hESC colony cultures. Optimized subprocesses were linked together to deliver a controlled complete process. We have demonstrated a rapid, high-throughput, and stable system for measurement of cell adherence and viability as robust markers of in-process and postrecovery cell state. We observed that measurement of adherence and viability of adhered cells at 1h postseeding was predictive of cell proliferative ability up to 96h in this system. Application of factorial design defined the operating spaces for cryopreservation and resuscitation, critically linking the performance of these two processes. Optimization of both processes resulted in enhanced reattachment and post-thaw viability, resulting in substantially greater recovery of cryopreserved, pluripotent cell colonies. This study demonstrates the importance of QbD concepts and tools for rapid, robust, and low-risk process design that can inform manufacturing controls and logistics.
Funding
The authors gratefully acknowledge the financial support from Pfizer Neusentis and the EPSRC centre for regenerative medicine.
History
School
- Aeronautical, Automotive, Chemical and Materials Engineering
Department
- Chemical Engineering
Published in
Tissue Engineering - Part C: MethodsVolume
20Issue
12Pages
941 - 950Citation
MITCHELL, P. ... et al., 2014. A Quality-by-Design approach to risk reduction and optimization for human embryonic stem cell cryopreservation processes. Tissue Engineering. Part C, Methods, 20 (12), pp.941-950.Publisher
© Mary Ann LiebertVersion
- AM (Accepted Manuscript)
Publisher statement
This work is made available according to the conditions of the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) licence. Full details of this licence are available at: https://creativecommons.org/licenses/by-nc-nd/4.0/Publication date
2014Notes
Final publication is available from Mary Ann Liebert, Inc., publishers: https://doi.org/10.1089/ten.tec.2013.0595.ISSN
1937-3384eISSN
1937-3392Publisher version
Language
- en