Assessing the toxic effects of DMSO on cord blood to determine exposure time limits and the optimum concentration for cryopreservation

作者：Fry L J^*; Querol S; Gomez S G; McArdle S; Rees R; Madrigal J A

来源：Vox Sanguinis, 2015, 109(2): 181-190.

DOI：10.1111/vox.12267

• cord blood
• cryopreservation
• dextran-40
• DMSO

Background and ObjectivesAdvantages of using cord blood (CB) over other sources of haematopoietic progenitor cells, such as bone marrow, include the ability to cryopreserve and bank the samples until requested for a transplant. Cryopreservation requires the addition of a cryoprotectant to prevent the formation of intracellular ice during freezing. Dimethyl sulphoxide (DMSO) is commonly used at a concentration of 10% (v/v); however, there is evidence to suggest this chemical is toxic to cells as well as to patients after infusion.
Materials and MethodsThe toxic effects of DMSO were assessed through cell viability and in vitro functional assays in fresh and post-thaw CB samples before determining the maximum exposure time and optimal concentration for cryopreservation.
ResultsA dose-dependent toxicity of DMSO was observed in fresh samples with 40% removing all viable and functional haematopoietic progenitor cells (HPC). In fresh and post-thaw analysis, minimal toxic effect was observed when cryopreservation was delayed for up to 1h after 10% DMSO addition. After thawing, DMSO washout was superior to dilution or unmanipulated when maintained for long periods (advantage observed 1h after thawing). Finally, the optimum concentration for cryopreserving CB was found to be 75 to 10% with detrimental effects observed outside of this range.
ConclusionThese results support the use of 75-10% as the optimal DMSO concentration and the maximum exposure time should be limited to <1h prior to freezing and 30min post-thaw.

• 出版日期2015-8