Abstract
Though in vivo animal models are commonly used in cancer biology and drug research, their imprecision and lack of physiological relevance pose a major hurdle for clinical translation. ATCC’s luciferase-labeled cell lines provide a simple, sensitive, and versatile means to measure specific biological processes through bioluminescence imaging. Because these engineered cell lines are a powerful tool for oncology and immuno-oncology research, it is important to consider the impact that low-temperature storage will have on the luciferase reporter functionality.
Cryopreservation using a controlled-rate freezer followed by storage in the vapor phase of liquid nitrogen is the gold standard for long-term preservation of mammalian cells at ATCC. It is a well-known phenomenon that cryopreservation and subsequent thawing are stressful processes that generally require a recovery period in culture for growth, metabolism, and functionality to return to normal. Because of this, it is important to understand how these processes impact the resulting recovery time required for our luciferase reporter cells to be able to provide the accurate and repeatable results expected.
In this study, we compared the immediate post-thaw viability, growth, and functionality characteristics of THP-1 luciferase reporter cells (ATCC TIB-202-NFkB-LUC2) to those of non-frozen controls. As expected, the cells immediately recovering from cryopreservation had different characteristics than those in fresh culture. Specifically, we identified an atypical cellular response to immediate post-thaw lipopolysaccharide (LPS) stimulation.
Observing the impact of preservation on the cell characteristics and understanding the mechanisms that lead to these phenotypic changes provide us with an opportunity to optimize our cryopreservation formulations and procedures toward reducing stress and enhancing recovery. Minimizing this recovery time will enable our customers to start their research faster.
Download the presentation to learn how cryopreservation affects luciferase reporter functionality
DownloadPresenters
Quinn Osgood, BSE
Lead Biologist, BioNexus Cryobiology, ATCC
Quinn specializes in preservation sciences with 10 years of experience in preservation related research. Quinn received dual BSE in BioEngineering and Mathematics from the University of Michigan. Prior to joining ATCC, Quinn founded a start-up and acquired NSF SBIR funding for the development of a novel dry-state preservation technology and advised a separate start-up on the development of a serum and DMSO free cryopreservation medium for cord blood preservation. At ATCC, Quinn has worked on enabling new product lines through the development of preservation capabilities in both cell biology and microbiology. Quinn also advises on modernization of preservation equipment set up and operation, and he aids in troubleshooting preservation related problems in both R&D and Production.
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.
