3D Hepatocyte Spheroids In Vitro Induction Screening

Human Primary 3D hepatocyte spheroids System
CYP1A2, CYP2B6, CYP3A4 mRNA expression
14-day data turnaround
72 hours Repeat-dose
$ 1K per compound*

Benefits Using 3D Hepatocyte Spheroids Culture System 

• Reduce risk of false positives 

• More closely mimic normal liver tissue micro-anatomy and function

• Maintains physiologically-relevant uptake and efflux hepatic transporters activity

• Higher baseline level of CYP450 

• Maintains physiologically-relevant intracellular concentrations of test compound 

• Allow you to interrogate mechanisms of action of your compound

• Better in vitro – in vivo correlations

• Cost-efficient and screening of higher number of compounds

 Cytochrome P450 (CYP) induction by a new chemical entity results in the increased amount and activity of relevant drug-metabolizing CYPs, which lead to reduce plasma levels of co-medicated drugs, resulting in a decrease in efficacy. Induction of cytochrome P450 enzymes can also lead to toxicity by increasing reactive metabolite formation. Because CYP induction could pose a significant risk to patients, induction-mediated drug-drug interaction (DDI) needs to be carefully evaluated to determine and/or predict their safety risk. The newly FDA guidance is a strong suggestion that the in vitro DDI studies should be done earlier in the drug discovery.

CYP induction may occur by stabilization of the mRNA or enzyme, decreasing the rate of CYP proteins degradation and nuclear receptor activation. The in vitro models will need to be carefully designed to ensure the experiments are physiologically relevant.

At MB Biosciences, we use human 3D hepatocyte spheroids culture for CYP450 enzyme induction screening assays. Our assay ensures that 3D environment that replicates native tissue, uptake and efflux hepatic transporters are functioning reflect physiologic conditions. We have demonstrated that our model has better in vitro – in vivo correlations and is superior at predicting clinical outcomes.

As a part of our CYP induction services, the CYP induction screening assays are designed for cost-efficient and screening of higher number of compounds. The 3D hepatocyte spheroids induction assays are available for CYP1A2, CYP2B6 and CYP3A4. Assays for other CYPs, UGTs and transporters are available upon request.

3D hepatocyte spheroids Cytochrome P450 induction Screening

Cytochrome P450 Induction Screening Protocol

MB Biosciences is the most advanced contract research organization offering CYP induction assay through a combination of our expertise in 3D cell culture models and extensive experience in CYP induction assessment.

Hepatocytes in 3D spheroid cultures were shown to build extensive bile canaliculi network throughout the spheroid represent a responsive and physiologically relevant in vitro liver model that maintains stable function over 5 weeks and is therefore well suited for repeated-dose testing

3D Spheroids showed a significantly higher baseline level of CYP450 activity and induction over monolayers. Positive staining in spheroids for albumin, BSEP and multidrug resistance-associated protein (MRP2) indicates the preservation of hepatocyte function within spheroids 3D environment that replicates native tissue. 

The CYP inducing potential of CYP2C19 mRNA in 2D culture was not stable. This is consistent with previous reports. Our results indicated that the inducibility of the CYP2C19 mRNAs in Human 3D primary hepatocyte spheroids culture relatively high and stable.

Four lots of cryopreserved hepatocytes were cultured as 2D format and 3D hepatocyte spheroids. After they were treated once daily for up to three consecutive days (72 hrs) with a vehicle control (0.1% DMSO), phenobarbital (1000 µM), rifampin (25 µM), troglitazone (20 uM) and Nifedipine (250 uM) with media replacement every 24 hrs, as appropriate. CYP450 2C19 mRNA expression was evaluated at 72 hrs by RT-PCR.

MB Biosciences provides a full range of products and services based on our three-dimensional (3D) cell culture platform. With the well-established 3D cell culture systems, our services include, but are not limited to, establishment and optimization of experimental protocols, a series of in vitro analysis and screening to improve in vivo relevancy.