
Types of hepatocytes: A comprehensive guide for researchers
Types of hepatocytes: A comprehensive guide for researchers
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Hepatocytes, the primary functional cells of the liver, are essential for various applications in research, including drug metabolism, toxicology, and disease modeling. Knowing the different types of available hepatocytes for research based on their origin, attachment capacity, and shipment method, is essential to selecting the most suitable model for accurate and efficient experimental outcomes.
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Classification by origin organism
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Human hepatocytes
Human hepatocytes are derived from human liver tissue and represent one of the most clinically relevant types of hepatocytes. Their genetic and functional alignment with human physiology makes them ideal for studying human-specific drug metabolism, liver diseases, and toxicity responses.
These cells are commonly used in pharmaceutical research to predict how drugs will behave in the human body, providing insights that animal models cannot fully replicate.
BeCytes is a first-class provider of primary human hepatocytes in diverse formats. Our cells are fully characterized, including the number of cells and viability per vial, cell morphology, attachment efficiency, and metabolization and induction capacity of cytochrome P450.
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Animal hepatocytes
Animal hepatocytes are isolated from non-human species, such as rats, mice, or dogs.
Primary animal hepatocytes and subcellular liver fractions (microsomes, cytosol, and S9 fraction) are model systems often used for a variety of in vitro screenings in preclinical research before advancing to clinical studies in human and animal health as well as environmental toxicology.
While they provide valuable data, interspecies differences must be considered in metabolic pathways when interpreting the results.
At BeCytes, we grant access not only to human liver cells but also to animal cells from diverse species, including beagle, cynomolgus monkey, minipig, horse, sheep, chicken, duck, several fishes, and rodents.
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Classification by state
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Primary Hepatocytes
Primary hepatocytes are freshly isolated from liver tissue, maintaining genotypic and phenotypic stability that closely mirrors in vivo conditions. They are considered the gold standard in liver research due to their ability to retain essential metabolic functions, such as cytochrome P450 enzyme activity, albumin production, and urea synthesis.
However, primary hepatocyte finite lifespan in culture poses a challenge, limiting their utility to short-term studies. Advances in cryopreservation and plating culture techniques have improved accessibility and functionality, allowing researchers to utilize primary hepatocytes in various experimental settings. Despite these challenges, their unparalleled physiological relevance ensures their prominence in preclinical research.
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Immortalized Hepatocytes
Immortalized hepatocytes or hepatocyte cell lines are genetically modified cells that proliferate indefinitely, making them ideal for high-throughput drug screening and gene-editing applications.
They provide a cost-effective and scalable alternative for long-term studies. However, immortalized hepatocytes often lack the full physiological relevance of primary hepatocytes, with limitations in drug-metabolizing enzymes and transporter expression. Compared to the immortalized cell line HepG2, differentiated lines, such as HepaRG, offer improved functionality but require complex differentiation protocols.
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Classification by attachment capacity
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Plateable hepatocytes
Plateable hepatocytes are a subset specifically prepared for adherence to culture surfaces. They are handy for long-term in vitro culture, making them ideal for drug toxicity testing, drug screening, and hepatocyte-based disease modeling applications.
While two-dimensional (2D) culture conditions allow these cells to adhere and form monolayers, this environment may cause a partial loss of metabolic functions and phenotypic characteristics, as it does not fully replicate the behavior of hepatocytes within the liver.
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3D Hepatocyte Culture
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Advancements in 3D cell culture techniques have revolutionized hepatocyte research, allowing for more physiologically relevant models that mimic the liver’s in vivo environment. Unlike traditional 2D cultures, 3D systems enable hepatocytes to maintain their native morphology, enhancing cell-to-cell interactions and preserving crucial liver-specific functions. These techniques often involve scaffolds, spheroids, organoids, or organ-on-chip, which recreate the liver niche and support the long-term viability of hepatocytes. By integrating biomaterials and co-culturing with non-parenchymal cells, 3D cell culture techniques are the most appropriate cell culture method to ensure hepatocytes’ full functionality in vitro.
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Suspension hepatocytes
Suspension hepatocytes are prepared for use in non-adherent conditions. These cells are optimized for short-term experiments where hepatocyte functionality is critical, such as metabolic clearance assays and enzyme activity studies.
Studies have shown that cytochrome P450 (CYP)-dependent activities are more stable in suspension hepatocytes during the first six hours of incubation compared to plateable hepatocytes. This stability is attributed to the cells’ ability to retain a more native three-dimensional (3D structure, which preserves higher metabolic activity.
One of their main advantages lies in their readiness. Suspension hepatocyte culture does not require cell plating or incubation to wait for attachment. However, their use is time-sensitive; suspension hepatocytes typically survive for about 4–6 hours, limiting their suitability for long-term studies.
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Classification by shipping method
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Cryopreserved hepatocytes
Cryopreserved hepatocytes are frozen using cryogenic techniques for long-term storage.
This preparation allows access to functional hepatocytes on demand, reducing variability across experiments and enabling standardized workflows, cryopreserved hepatocytes retain critical metabolic activities. The main advantage of cryopreserved hepatocytes is that the samples can be delivered, cryopreserved, and stored, allowing them to be used whenever it is most convenient.
They are usually shipped using dry ice or the vapor phase of liquid nitrogen contained in a dewar to maintain the temperature between -140 °C and -160 °C.
Our certificate of analysis includes post-thaw lot information, such as post-thaw viability, recommended seeding density, and cell morphology. Find more info about our cryopreserved hepatocytes here.
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Fresh hepatocytes
Fresh hepatocytes are liver cells that are immediately isolated and used without undergoing any cryopreservation process. These hepatocytes maintain optimal metabolic activity and cellular viability, making them highly valuable for studies requiring precise liver function replication.
After isolation, fresh hepatocytes can be received either in suspension or as plated cells. At BeCytes, our extensive network of hospitals and laboratories can provide fresh hepatocytes to Europe.
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Plated hepatocytes
Plated hepatocytes offer a tailored solution for researchers seeking specific experimental setups. Researcher can select the donor, and the hepatocytes are thawed, plated, and prepared in a controlled environment to ensure optimal viability and functionality. The plating process allows researchers to obtain detailed data on cell plating and performance, making it possible to conduct experiments under highly reproducible conditions. Once prepared, the plated hepatocytes are delivered directly to the laboratory, ready for use.
For a complete description of our products and services, do not hesitate to check our inventory.
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References
Blanchard N, Richert L, Notter B, et al. Impact of serum on clearance predictions obtained from suspensions and primary cultures of rat hepatocytes. European Journal of Pharmaceutical Sciences. 2004;23(2):189-199. doi:10.1016/j.ejps.2004.07.007
Guo L, Dial S, Shi L, Branham W, Liu J, Fang JL, et al. Similarities and differences in the expression of drug-metabolizing enzymes between human hepatic cell lines and primary human hepatocytes. Drug Metab Dispos. 2011 Mar;39(3):528–38. doi: 10.1124/dmd.110.035873
Jouin D, Blanchard N, Alexandre E, et al. Cryopreserved human hepatocytes in suspension are a convenient high, throughput tool for the prediction of metabolic clearance. European Journal of Pharmaceutics and Biopharmaceutics. 2006;63(3):347-355. doi:10.1016/j.ejpb.2006.01.014
Rodríguez-Antona C, Donato MT, Boobis A, Edwards RJ, Watts PS, Castell JV, et al. Cytochrome P450 expression in human hepatocytes and hepatoma cell lines: molecular mechanisms that determine lower expression in cultured cells. Xenobiotica. 2002 Jun;32(6):505–20. doi: 10.1080/00498250210128675
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