Bile Acid-Dependent Hepatotoxicity Evaluation by Human Hepatocytes

Bile acid (BA) accumulation in the liver causes hepatocyte inflammation, necrosis, or apoptosis, which is one of the fundamental mechanisms of cholestatic drug-induced liver injury (DILI). Creative Biolabs launches a sandwich-cultured human hepatocytes (SCHHs) model for the assessment of your drug’s bile acid-dependent hepatotoxicity

Introduction of Bile Acid-Dependent Hepatotoxicity

Hepatocellular damage and cholestasis can be caused by a variety of reasons, including mitochondrial malfunction, cytotoxicity, the production of reactive metabolites, and the bile salt export pump (BSEP) inhibition.

BSEP is located on the apical side of the hepatocyte plasma membrane, where it is important in the excretion of bile acids (BAs) from the liver. BSEP dysfunction due to mutations in several genes leads to severe intracellular accumulation of BAs within the liver.

Bile Acid-Dependent Hepatotoxicity Evaluation by Sandwich-cultured Human Hepatocytes

Construction Method

Hepatocyte isolation before establishing sandwich-culture hepatocytes is frequently isolated from both animal and human livers by a two-step collagenase perfusion. First, the liver tissue is perfused with a calcium-free buffer solution containing a cation-chelating agent, leading to the removal of blood and the disruption of cell-cell contacts. In the second step, perfusion is continued with a buffer containing calcium and collagenase to digest the surrounding tissue. Finally, XenoTech Optical Medium, Matrigel Overlay, and Bile Acid Mix are added to the 96-well plate to mimic in vivo conditions.

Test Principle

Drugs with BSEP inhibitory function are expected to exhibit BA accumulation in hepatocytes. As this state persists, hepatocytes rupture to release lactate dehydrogenase (LDH) and exhibit hepatotoxicity.

Test Procedure

In the presence of BAs, SCHHs were exposed to each drug for 24 hours at non-toxic drug and BA concentrations. If the test drug and/or its metabolites prevent bile acid efflux from hepatocytes, bile acid buildup leads to cell death (apoptosis), which can be identified by the release of LDH into the culture medium.

Advantages

1. The development of complete canalicular networks and polarized excretory function.
2. Improved hepatocyte morphology and viability, as well as normal secretion of various liver-specific proteins and chemical molecules such as urea, albumin, and bile acids.
3. Promotion of functional bile duct networks and development of gap junctions over days in culture.
4. Maintenance of cell polarity and the expression of transporters and metabolic enzymes.

Applications

• Reflect on the impact of medication metabolism on BA-dependent DILI.
• Determination of the cytotoxicity of BAs and other chemicals.
• Estimation of BA-dependent drug-induced cytotoxicity.
• Effect of CYP450 metabolism on BA-dependent toxicity.
• Detection of medication candidates with cholestatic signature.

This hepatocyte-based in vitro assay offers a special tool for the early and accurate identification of medication candidates with cholestasis risk. If you need an evaluation of bile acid-dependent hepatotoxicity, please feel free to contact us. We are here to help and provide the support you need.

Related Sections:

• 3D Biology Based Ocular Toxicity Evaluation Services
• 3D Biology Based Skin Toxicity Evaluation Services
• 3D Biology Based Cardiotoxicity Evaluation Services
• 3D Biology Based Hepatotoxicity Evaluation Services
• 3D Biology-Based Neurotoxicity Evaluation Services
• 3D Biology-Based Nephrotoxicity Evaluation Services
• 3D Biology-Based Gastrointestinal Toxicity Evaluation Services
• 3D Biology Based Phototoxicity & Photoallergy Evaluation Services
• 3D Biology Based Pancreatic Endocrine Toxicity Evaluation Service
• 3D Biology Based Inhalation Toxicity Evaluation Services
• 3D Biology Based Oral Irritation Evaluation Services