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The Need for Standardized Cryoprocessing on Preservation Outcomes

Large cryopreservation storage tanks at biorepository.

ISBER 2023 Annual Meeting & Exhibits

Virtual exhibit

June 06, 2023



Recent literature indicates that the characteristics of ice formation during cryopreservation can impact preservation outcomes in eukaryotic cells. Ice formation can be modulated by various chemical approaches such as anti-freeze proteins or reagents and other physical means such as high temperature ice nucleation and controlled rate freezing. Certain non-adherent human cell lines have poor post-cryopreservation outcomes characterized by significant loss of viability 24 hours post-thaw, which could be due to variation of ice formation characteristics during cryoprocessing. It is a common practice to use passive freezing containers for small scale cryopreservation. However, such freezing conditions have limited scalability and offer very little control over the freezing process. The current work highlights the need for standardization in freezing processes in cryopreserving biomaterials in production environments.


In this study, preservation outcomes of THP-1 human monocytes isolated from a monocytic leukemia patient (ATCC TIB-202) were compared from multiple controlled rate freezing (CRF) temperature profiles using an identical cryopreservation medium. Cell recovery was measured using standard viability, growth, and metabolic activity. Finally, the thermal characteristics of each CRF profile were analyzed using differential scanning calorimetry.


All recovery metrics recorded were moderately lower when compared to the non-frozen control. Additionally, there was statistically no difference in viability and metabolic activity between the different freezing methods. However, there was a reduction in the variability between the CRF profiles when compared to the passive freezing container. We also report differences in the thermal characteristics between the different freezing profiles.


This work reconfirms that batch-to-batch variability remains a concern in cryopreservation outcome of mammalian cells; standardization of CRF profile could be an important strategy to address the issue. Further investigation is required to identify the exact mechanisms of variability reduction during cryoprocessing.

Download the poster to learn about need for the standardization of freezing processes in cryopreservation.



Lukas Underwood.jpg

Lukas Underwood, PhD

Scientist, BioNexus Cryobiology, ATCC

Dr. Underwood specializes in mammalian cell preservation and characterization. He has a PhD in Mechanical Engineering and Applied Sciences from the University of Michigan with a focus in biological engineering. Lukas has extensive experience in and spectrographic and thermodynamic characterization of mammalian cell preservation and formulation development.

Dr. Underwood joined ATCC is March 2022 and is working on the development of novel preservation formats for Cell Biology products. Since joining ATCC, Dr. Underwood has collaborated with and consulted for several of the Cell Biology R&D groups located on the Gaithersburg campus.