Abstract
Cell culture, a mainstay in biological research, faces challenges like prolonged culture times, high labor, and resource demands, leading to inefficiencies and risks such as genetic drift and contamination. Cryopreservation is often employed alongside cell culture, facilitating storage and on-demand utilization of cells, thereby reducing the continuous need for active cultures. Yet, cryopreservation itself presents unique challenges, including significant cell loss and requisite stabilization periods for even the most robust cell lines. In response to these challenges, innovative cryoprotectants coupled with optimized processes have led to the development of 'assay-ready' cells. These cells represent a significant leap forward; they revive quickly after thawing in a format that further diminishes cell culture requirement and are primed for immediate assay integration. This innovation streamlines cell applications, offering a practical solution to the inefficiencies of traditional cell culture practices.
In this study, THP-1 (ATCC TIB-202) and HepG2 (ATCC HB-8065) cell lines were selected for their significant application in biological research and distinct culture requirements, offering a broad perspective on assay-ready cell efficacy. Immediately following cryopreservation and thawing, cells were assessed against a continuous culture control. Viability and growth metrics were obtained using an automated cell counter. For THP-1, LPS-induced NFkB activation was measured with a luminescent reporter assay, while HepG2 was evaluated for post-thaw attachment efficiency and acetaminophen dose-response.
The results demonstrated that both THP-1 and HepG2 cell lines maintained over 90% viability post-thaw. Despite minimal differences in general recovery, the LPS response in THP-1 and acetaminophen response in HepG2 showed strong agreement with the positive control. These findings underscore the potential for assay-ready application of these cell lines. However, comprehensive validation in specific assay conditions is required for mainstream applications. Laboratory professionals are thus encouraged to consider the integration of assay-ready cells into their workflows, to save time and resources in assay workflows.
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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.
