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|Title: ||A Quality-by-Design approach to risk reduction and optimization for human embryonic stem cell cryopreservation processes|
|Authors: ||Mitchell, Peter D.|
Hourd, Paul C.
Williams, David J.
Thomas, Robert James
|Keywords: ||Stem cells|
|Issue Date: ||2014|
|Publisher: ||© Mary Ann Liebert|
|Citation: ||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.|
|Abstract: ||It 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.|
|Description: ||Final publication is available from Mary Ann Liebert, Inc., publishers: https://doi.org/10.1089/ten.tec.2013.0595.|
|Sponsor: ||The authors gratefully acknowledge the financial support from Pfizer Neusentis and the
EPSRC centre for regenerative medicine.|
|Version: ||Accepted for publication|
|Publisher Link: ||http://dx.doi.org/10.1089/ten.tec.2013.0595|
|Appears in Collections:||Published Articles (Chemical Engineering)|
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