Why is the use of 3D models in hepatic research becoming more common?
Over the last decade many new in vitro models have been established and optimized for 3D studies of hepatic systems. A few examples include sandwich cultures, chip systems, bioprinting and stem cell–derived organoids and primary cell–based spheroids and organoids. Hepatic spheroids demonstrate physiologically relevant features such as increased cell-to-cell interactions, superior longevity and enhanced hepatocyte-specific function. As a result, spheroid-qualified hepatocytes have recently emerged as a tool facilitating creation of human liver models. Start researching in the third dimension today.
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Why use Gibco Spheroid-Qualified Hepatocytes?
- Improved culture longevity—up to 21 days, that’s more than 4x longer than traditional 2D monolayer culture
- Design your own schedule—cells are ready for plating when you are
- Interrogate pathways of interest—applicable for toxicity, metabolism, or disease modeling
The 3D advantage in hepatic research
In addition to the longevity benefit, Gibco hepatocytes maintained as spheroids demonstrate superior function, as demonstrated by increases in:
- Albumin protein production
- CYP3A4 enzyme activity
- CYP3A4 gene expression
- Albumin gene expression
Researchers have found that hepatocytes cultured in 3D are more sensitive to toxic compounds such as rosuvastatin and troglitazone.1 A separate publication reported that primary human hepatic spheroids were an effective model for studying liver fibrosis.2
Simplified workflow for Gibco Spheroid-Qualified Hepatocytes
Figure 1. Workflow of assembly and characterization of primary hepatocytes in 3D spheroid culture. (A) Spheroids were imaged in phase at 10x magnification. These images show spheroid formation by day 5 of culture. To learn more about Gibco Human Spheroid-Qualified Hepatocytes, see the product page that includes the user guide.
Have questions or need help getting started? Contact us.
If you’d like a demonstration of Gibco Cryopreserved Spheroid-Qualified Hepatocytes from one of our experienced scientists, follow the link below to complete the contact form. You can also complete the contact form to request information from our scientists about using cells such as HepaRG, and animal hepatocytes to create 3D models. Contact us
Albumin production is upregulated in hepatic spheroids
Figure 2. Albumin secretion in 2D and 3D spheroid hepatic cultures. The concentration of albumin secreted is normalized to the total number of cells per well.
CYP3A4 activity increases hepatic spheroids
Figure 3. CYP3A4 activity in 2D and 3D spheroid hepatic cultures. CYP3A4 activity was measured using a P450-Glo CYP3A4 Assay with Luciferin-IPA. CYP3A4 activity was found to be significantly higher in the 3D spheroids than in the 2D culture. The data presented are the mean ± SEM (n = 3 for the 2D culture, n = 8 for the 3D spheroids).
Step-by-step application notes
Our Gibco 3D spheroid–qualified human hepatocytes have been characterized to show stable morphology, viability, and hepatocyte-specific functions for at least 3 weeks. Learn more about how to get started today with our Hepatic growth and liver function guide and app note
Review our most recent posters on hepatocyte spheroids from ISSX and SOT
Introducing the Gibco Hepatic Spheroid Kit
Our Gibco Hepatic Spheroid Kit is a comprehensive system for the generation of longer-lived hepatic spheroids, within five days after a single hands-on session of 60 minutes or less.
Critical components of the Gibco Hepatic Spheroid Kit include:
- Gibco Human Spheroid-Qualified Hepatocytes—off-the-shelf cryopreserved human cells that allow you to culture and generate hepatic spheroids in your own lab. Each vial contains >5 million viable cells, sufficient for at least sixteen 96-well plates of hepatic spheroids.
- Thermo Scientific Nunclon Sphera 96-well U-bottom Microplate—with its super-low cell attachment surface, consistently form hepatic spheroids for disease modeling and drug screening.
- Optimized Gibco media, matrices and supplements—the most cited brand in 3D publications for organoids, spheroids and 3D models*
*Based on a third-party market report covering papers cited for disease modeling of organoids and spheroids with primary or stem cells as the starting cell type.
Human hepatic stellate cells
Human hepatic stellate cells (HSC) are the major contributor to collagen deposition following liver damage, where hepatic stellate cells activate to a phenotype characterized by increased proliferation, motility, contractility, and synthesis of extracellular matrix components that result in progressive liver fibrosis. Activated HSCs are ideal for building hepatic co-cultures with primary hepatocytes to for in vitro study of diseases such as fibrosis.
***The cells are obtained from a single donor and provided in a cryopreserved format identical to single donor plateable hepatocytes
Important! For best results, review our application note on 3D Hepatic culture and liver function, using the Gibco and Thermo Fisher Scientific suite of products. Hepatocyte lots have been qualified in Nunclon Sphera plates only, and use of other plates can lead to variability in spheroid formation.
- Proctor WR, Foster AJ, Vogt J, et al. (2017) Utility of spherical human liver microtissues for prediction of clinical drug-induced liver injury. Arch Toxicol 91: 2849-2963. DOI: 10.1007/s00204-017-2002-1
- Kozyra M, Johansson I, Nordling A, et al. (2018) Human hepatic 3D spheroids as a model for steatosis and insulin resistance. Sci Rep 8:14297. DOI: 10.1038/s41598-018-32722-6
We offer one-on-one training with the help of our Field Application Scientists (FAS team) from basic start-up questions and demos up-to getting started in 3D for ADME/DMPK research. reach out today to speak to our team of experts.
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