The effect of Me$_{2}$SO overexposure during cryopreservation on HOS TE85 and hMSC viability, growth and quality
Publication Date
2016-12-01Journal Title
Cryobiology
ISSN
0011-2240
Publisher
Elsevier
Volume
73
Issue
3
Pages
367-375
Language
English
Type
Article
This Version
VoR
Metadata
Show full item recordCitation
Morris, T., Picken, A., Sharp, D., Slater, N., Hewitt, C., & Coopman, K. (2016). The effect of Me$_{2}$SO overexposure during cryopreservation on HOS TE85 and hMSC viability, growth and quality. Cryobiology, 73 (3), 367-375. https://doi.org/10.1016/j.cryobiol.2016.09.004
Abstract
With the cell therapy industry continuing to grow, the ability to preserve clinical grade cells, including mesenchymal stem cells (MSCs), whilst retaining cell viability and function remains critical for the generation of off-the-shelf therapies. Cryopreservation of MSCs, using slow freezing, is an established process at lab scale. However, the cytotoxicity of cryoprotectants, like Me$_{2}$SO, raises questions about the impact of prolonged cell exposure to cryoprotectant at temperatures >0 °C during processing of large cell batches for allogenic therapies prior to rapid cooling in a controlled rate freezer or in the clinic prior to administration. Here we show that exposure of human bone marrow derived MSCs to Me$_{2}$SO for ≥1 h before freezing, or after thawing, degrades membrane integrity, short-term cell attachment efficiency and alters cell immunophenotype. After 2 h's exposure to Me$_{2}$SO at 37 °C post-thaw, membrane integrity dropped to ∼70% and only ∼50% of cells retained the ability to adhere to tissue culture plastic. Furthermore, only 70% of the recovered MSCs retained an immunophenotype consistent with the ISCT minimal criteria after exposure. We also saw a similar loss of membrane integrity and attachment efficiency after exposing osteoblast (HOS TE85) cells to Me$_{2}$SO before, and after, cryopreservation. Overall, these results show that freezing medium exposure is a critical determinant of product quality as process scale increases. Defining and reporting cell sensitivity to freezing medium exposure, both before and after cryopreservation, enables a fair judgement of how scalable a particular cryopreservation process can be, and consequently whether the therapy has commercial feasibility.
Keywords
human mesenchymal stem cells, HOS TE85, cryopreservation, dimethylsulfoxide, bioprocessing, toxicity
Sponsorship
The authors would like to acknowledge the Engineering and Physical Sciences Research Council (EPSRC; UK, EP/F500491/1) and Bioprocessing Research Industry Club (BBSRC/BRIC; UK, BB/I017602/1) for their support and funding.
Funder references
Biotechnology and Biological Sciences Research Council (BB/I016961/1)
Embargo Lift Date
2100-01-01
Identifiers
External DOI: https://doi.org/10.1016/j.cryobiol.2016.09.004
This record's URL: https://www.repository.cam.ac.uk/handle/1810/262708
Rights
Attribution 4.0 International, Attribution 4.0 International, Attribution 4.0 International, Attribution 4.0 International
Statistics
Total file downloads (since January 2020). For more information on metrics see the
IRUS guide.