HepatoXcell™: Reliable, High-Quality Primary Human Hepatocytes for Breakthrough Discoveries

What are primary human hepatocytes?

Primary human hepatocytes (PHHs), which constitute approximately 80% of the liver's mass, are distinctive due to their ability to possess multiple nuclei within a single cell, a result of their rapid division. The liver possesses the remarkable ability to regenerate to its original size following injury or surgical resection. This regenerative capacity ensures that—even after significant tissue damage or removal—the liver will eventually restore itself to its initial dimensions.³ However, once PHHs are extracted from the liver, they cease to divide, a phenomenon that remains under investigation by researchers. Consequently, the number of PHHs obtained during isolation is finite and cannot be expanded. When cultured in a sandwich configuration, PHHs exhibit a finite lifespan of about one week (provided they are of high quality) and typically display a cobblestone morphology.⁴ These inherent limitations, coupled with their sensitivity, render PHHs challenging to isolate and maintain. As a result, researchers continuously seek stable sources of PHHs for applications in toxicology and drug discovery.

How are PHHs obtained and characterized?

Human livers can be obtained either as surgical discards from living donors or as whole livers procured through organ procurement organizations, which work with hospitals to acquire liver tissue from deceased donors. These whole liver tissues may be diseased or otherwise deemed unsuitable for transplantation. Rigorous screening for multiple diseases (e.g., HIV, HPV, HCV) is conducted, and the tissues are carefully stored, meticulously packaged, and shipped prior to isolation. During the isolation process, the tissue undergoes enzymatic digestion to separate the cells from the connective tissue. A purification step is then performed to assess cell viability and yield.⁵

Post-isolation, PHHs can be either plated fresh or cryopreserved in vials. While fresh PHHs are typically higher quality hepatocytes, their limited lifespan necessitates the use of cryopreserved PHHs for prolonged storage and subsequent utilization. The limited availability of liver tissue and the complexity of isolation underscore the importance of cryopreserved PHHs. The cryopreservation process involves optimizing a cryopreservation solution and employing a controlled-rate freezer, which utilizes liquid nitrogen (LN₂) to cool the cells at a user-defined rate.⁶ Hepatocytes are generally regarded as sensitive cell types that are prone to cell death following isolation due to the absence of connective tissues and cell-cell interactions, which are essential for optimal cellular functionality.⁷

Hepatocyte cell culture is commonly conducted using a sandwich culture technique, wherein the cells are plated on collagen-coated plates and subsequently overlaid with an extracellular matrix.⁸ This method facilitates the recreation of the native connective tissue environment, thereby maintaining high plateable quality of the cells. However, the plateable quality of hepatocytes still varies between batches.⁷ While some batches can achieve the characteristic polygonal, cobblestone-like morphology with nearly 100% cellular confluence, others fail to plate well and are more suitable for suspension culture. Although batch-to-batch variations in the quality of PHHs are beyond control, early characterization of the hepatocytes is beneficial. Such characterization aids in categorizing the PHHs into plateable versus suspension lots,⁹ or even identifying their suitability for other formats such as advanced three-dimensional (3-D) culture.¹⁰ Characterization of PHHs include evaluating post-thaw viability, plateability, metabolic activity, mRNA expression, and transporter activity. This characterization is essential for assessing the suitability of a batch for plateable or suspension culture, as well as for 3-D culture applications.¹¹

What are the functions and benefits of hepatocytes? 

In the field of toxicology, hepatocytes are employed for diverse modeling and testing applications, including absorption, distribution, metabolism, and excretion-toxicology (ADME-TOX) studies; drug screenings; liver disease modeling; and advanced 3-D modeling, such as microphysiological systems (MPS) applications. During the investigation of novel pharmaceuticals or examining the hepatic effects of established drugs, ADME-TOX assays are indispensable. These assays provide critical insights into the absorption, distribution, metabolism, excretion, and toxicity of investigational new drugs (INDs).¹² Additionally, hepatocytes are utilized to model liver diseases, including fatty liver, alcoholic liver disease, metabolic disorders,¹³ hepatitis, cirrhosis, and liver cancer. The utilization of PHHs significantly enhances the physiological relevance of these disease models. Furthermore, the employment of PHHs assists researchers in adhering to the principles of the 3Rs, which seek to replace, reduce, and refine the use of animal models in scientific research. Usage of PHHs not only eases the shift from traditional 2-D cultures to advanced cellular models but also creates physiologically relevant in vitro systems, which can greatly enhance the potential for groundbreaking advancements in drug discovery and toxicology testing.¹⁴

Where can you source reliable hepatocytes to take your research to the next level?

Look no further than ATCC! With the launch of HepatoXcell™, researchers now have access to PHHs that are supported by ATCC’s renowned legacy of quality assurance and credibility. ATCC's new HepatoXcell™ products include three distinct formats of PHHs along with three associated media: HepatoXcell™ Pro (ATCC® PCS-450-011™), which guarantees viable cells for at least 7 days in 2-D sandwich culture, HepatoXcell™ Plus (ATCC® PCS-450-010™), which ensures plateable cells for a minimum of 3 days, and HepatoXcell™ Eco (ATCC® PCS-450-012™), which is perfect for suspension culture. The HepatoXcell™ product line also features three specialty media designed to optimally transition PHHs from a cryopreserved to cultured state: HepatoXcell™ Thawing Medium (ATCC® PCS-450-032™), HepatoXcell™ Plating Medium (ATCC® PCS-450-034™), and HepatoXcell™ Maintenance Medium (ATCC® PCS-450-038™). To guarantee superior quality, HepatoXcell™ PHHs are shipped directly to customers in temperature-monitored liquid nitrogen vapor dewars. This meticulous shipping process ensures the integrity of the cryopreserved hepatocytes, which can be stored for many years without compromising their essential performance or functionality. To learn more about HepatoXcell™ products visit www.atcc.org/HepatoXcell.  

What makes ATCC's hepatocyte offering the best choice for your research needs?

For the past 100 years, ATCC has been dedicated to providing its customers with the highest quality assurance. The HepatoXcell™ products, isolated under ISO-compliant conditions, uphold the same exceptional quality assurance that ATCC has delivered. HepatoXcell™ hepatocytes offer numerous advantages, specifically designed to make hepatic research work easier and enhance efficiency. For the first time in the industry, ATCC is bringing a 7-day plateable format, HepatoXcell™ Pro, ensuring a reliable hepatocyte culture period to meet all ADME-Tox assay needs, including transporter and induction assays as well as studying the clearance of low turnover compounds. Additionally, the 3-day plateable HepatoXcell™ Plus and suspension-grade HepatoXcell™ Eco are perfect for assays that don't require long-term cell cultures. All these options are offered at highly competitive prices, commensurate with their respective qualities.

Researchers frequently face challenges in identifying the appropriate hepatocytes suitable for their workflow due to substantial variations in demographics, gene expression, enzyme activities, and other parameters across different lots. The HepatoXcell™ lot selection tool simplifies this process by providing instant access to the certificates of analysis for all lots, complete with detailed characterization information, at the click of a mouse. Each hepatocyte lot will be accompanied by transcriptomics data generated under ISO 9001–compliant conditions, which will be available to researchers for further analysis. This high-quality 'omics data will enable researchers to bypass the time- and resource-intensive genetic characterization step while facilitating the identification of data most relevant to their research.

With this introduction of high-quality PHHs supported by specialty media and lot selection informatics support, ATCC is looking forward to empowering researchers with innovative solutions for the utilization of advanced hepatocyte cultures in cutting-edge research. Our reliable and superior hepatocyte cells will significantly contribute to the advancement of hepatic research, paving the way for breakthroughs that enhance human health.

Did you know?

ATCC offers thawing medium, maintenance medium, and plating medium that support the growth and handling of HepatoXcell™ primary human hepatocytes.

References

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