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Accelerate Cell-Based Assays with the ThawReady™ THP-1 NF-κB-Luc2 Reporter Line

Poster
ELISA plate to measure OD with microplate reader. Microtiter plate (96 well) reader for biochemistry analysis.

Society for Laboratory Automation and Screening (SLAS) 2025 Annual Meeting

San Diego, California, United States

January 27, 2025

Abstract

Introduction

Cell-based assays are pivotal in the drug development process. However, the inherent variability of cultured cells, which may result from contamination or phenotypic drift associated with long-term cultivation, poses a significant challenge. In addition, the cultivation of cells itself also involves labor-intensive lab procedures that can impact the quality and functionality of the cells. Standardization of the assay protocols, reagents, and biological materials used in cell-based assays is therefore essential for achieving reliable assay performance. The limitations of maintaining continuous culture have led to a demand for readily available cells for immediate use in cell-based assays. Assay-ready cells (ARCs) meet this demand, offering significant advantages like time and resource savings, improved data quality, reduced batch-to-batch variation, and increased flexibility. 

Leveraging our proprietary animal by-product (ABP)-free cryopreservation medium and well-established cell culture practices, ATCC has developed a THP-1 NF-κB-Luc2 ARC product: ThawReady™ THP-1 NF-κB-Luc2 (ATCC TIB-202-NFkB-LUC2-AR). Here, we optimized the culture and freezing conditions to provide cells with high post-thaw viability and functionality. These ARCs can be used directly from liquid nitrogen storage by simply thawing and seeding into assay plates, thereby eliminating lengthy cell expansion processes and accelerating pharmaceutically active compound discovery, biocompatibility testing, and assay development. 

Methods

To develop the ThawReady™ THP-1 NF-κB-Luc2 cells, THP-1 NF-kB-Luc2 cells (ATCC TIB-202) were cultivated following ATCC’s thawing and culture processes. Cell viability was measured using a Vi-CELL® BLU cell viability analyzer (Beckman Coulter®). NF-κB reporter activity evaluated by using Bright-glo™ (Promega®) and a luminometer (Glomax™).

Results

ThawReady™ THP-1 NF-kB-Luc2 cells consistently deliver high post-thaw viability, exceeding 80% across three distinct batches, with negligible intra-lot and inter-lot variation. Our functional evaluations reveal the luciferase activity in ThawReady™ THP-1 NF-kB-Luc2 cells increase an impressive greater than 200-fold in response to LPS stimulation. The NF-κB reporter retains sensitivity even at eight hours post-thaw rest, supporting that the ThawReady™ THP-1 NF-κB-Luc2 cells are ready to use for assaying upon thawing and are suitable for a one-day workflow. In addition, NF-κB reporter activity in ThawReady™ THP-1 NF-kB-Luc2 cells displays a dose-dependent manner in response to NF-κB activators and inhibitors stimulation. 

Conclusion

Using proprietary cryopreservation media and highly standardized cell culture practices, ATCC has developed an easy-to-use, highly functional cell line to accelerate cell-based assays: ThawReady™ THP-1 NF-κB-Luc2 (ATCC TIB-202-NFkB-LUC2-AR). These cells can be used directly as a standardized "reagent" in cell-based assays, eliminating the need for additional culturing. Our cells consistently exhibit high post-thaw viability with minimal intra-lot and inter-lot variation and offer functionality comparable to the parental THP-1 NF-κB-Luc2 cells. Incorporating our highly sensitive and reliable ThawReady™ THP-1 NF-kB-Luc2 cells into experiments will simplify your research processes and offer exceptional consistency, convenience, and efficiency.


Download the poster to learn more about the development of the THP-1 NF-κB-Luc2 reporter line.

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Presenter

headshot of Rajnee Kanwal

Rajnee Kanwal, PhD

Scientist, Cell Biology R&D, ATCC

Dr. Rajnee Kanwal is scientist in the product development group of the R&D department at ATCC. Dr. Kanwal has extensive experience in biomedical research with her research career extended in molecular biology, cancer biology and epigenetics, drug discovery and development, and assay development and qualification. Prior to joining ATCC, Dr. Kanwal received her PhD in Biosciences from Jamia Millia University, India, where her research focused on functional genomics, drug discovery, and drug development. She completed her postdoctoral training from Case Western Reserve University in the areas of prostate and breast cancer, and subsequently she worked as a research scientist in Learner Research Institute, Cleveland Clinic, to develop 3-D organoid models for colon cancer.

ELISA plate to measure OD with microplate reader. Microtiter plate (96 well) reader for biochemistry analysis.

ThawReady™ by ATCC Logo

Cell-based assays have lengthy timelines due to the requirement of cell expansion processes to get a synchronized cell stock. To speed your timelines while providing you with the consistency you need, ATCC developed ThawReady™ Assay Ready Cells. ATCC ThawReady™ products will streamline your workflows by months, allowing you to focus on advancing drug discovery and development. You simply thaw, plate, and go.

Explore ThawReady™ Assay Ready Cells