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Development of the ThawReady™ THP-1 Product for Cell-Based Assays

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 28, 2025

Introduction

Cell-based assays are valuable tools for basic research and drug discovery. Traditionally, these assays involve thawing frozen cells and culturing them through multiple passages to reach the required density for the assay. However, in continuous culture, most cells do not maintain their characteristics indefinitely. As the passage number increases over time, cells may acquire unintended or unexpected characteristics, and their responsiveness to certain modulators can change—a phenomenon known as phenotypic drift. Phenotypic drift presents one of the greatest challenges in achieving consistent, high-performance cell-based in vitro assays. Additionally, traditional cell-based assays have several disadvantages such as the labor required to maintain cell culture, the cost of consumables such as media and laboratory plasticware, the need for laboratory space, and the use of dedicated equipment like incubators.

The disadvantages of maintaining continuous cultures have driven the demand for cell products that are ready for immediate use in assays. To meet this need, ATCC has developed an assay-ready cell (ARC) product using the THP-1 cell line: ThawReady™ THP-1 (ATCC TIB-202-AR). In this study, we showcase the development of this novel product and demonstrate that it consistently exhibits high post-thaw viability and can differentiate into macrophage-like cells that express the appropriate markers and display expected functional attributes.

Methods

To establish the cell expansion process for ThawReady™ THP-1 cells, we grew parental THP-1 cells (ATCC TIB-202) in RPMI-1640 (ATCC 30-2001) supplemented with 10% Fetal Bovine Serum (ATCC 30-2020) and 0.9 µL/mL of 2-Mercaptoethanol (Thermo Fisher Scientific® #21985-023) following ATCC cell culture protocols. A proprietary animal by-product (ABP)-free cryomedium was developed to freeze the ThawReady™ THP-1 cells. Cell viability was measured using a Vi-CELL® BLU cell viability analyzer (Beckman Coulter®). 

Results

ATCC TIB-202-AR cells consistently display high post-thaw viability; 3 batches tested in house exhibit viabilities > 80%. In addition, ATCC TIB-202-AR cells exhibit low intra-lot and inter-lot variation and demonstrate reproducibility in achieving optimal performance in cell-based assays. Following a macrophage differentiation protocol, ThawReady™ THP-1 cells can differentiate into macrophage-like cells that express the appropriate markers, including CD14 and CD36, and maintain key functional attributes such as phagocytic capacity.

Conclusions

Leveraging our proprietary ABP-free cryopreservation media and well-established high-standard cell culture practices, ATCC has developed a highly functional ThawReady™ THP-1 cell product. It consistently exhibits high post-thaw viability with low intra-lot and inter-lot variation and shows characteristics and functionality equivalent to the parental THP-1 cells. Our ThawReady™ THP-1 cells demonstrate consistency and reproducibility in achieving optimal performance in cell-based assays, offering advantages like long-term access to a consistent resource, more flexible scheduling, and cost savings. This novel product allows for extensive biopharmaceutical studies while avoiding the lengthy and costly development typically required for establishing cell-based assays.

 

Download the poster to learn more about the development of the ThawReady™ THP-1 product.

Download

Presenter

Diana Douglas, headshot.

Diana Douglas, BS

Lead Biologist, ATCC

Diana Douglas is a Lead Biologist at ATCC. For the last nine years, she has focused her research on the development of advanced biological models including the use of CRISPR/Cas9 gene-editing technology. Previously, Ms. Douglas worked at the Baker Institute for Animal Health at Cornell University and the Dalton Cardiovascular Research Center at the University of Missouri, where her research focused on the mechanisms of necrotic cell death in heart disease. Ms. Douglas attended Truman State University where she obtained a Bachelor of Science in Biology.

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