Human iPSC-derived hepatocytes

Functional, consistent in vitro models to study hepatotoxicity, drug metabolism and disease

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Hepatocytes account for over 80% of the liver's mass and are responsible for its main functions, including lipid and glucose metabolism, storage of macronutrients, secretion of plasma proteins, detoxification and xenobiotic metabolism. Given their extensive biological role, hepatocytes are a critical cell type for both toxicology research and studying debilitating genetic and life-threatening diseases, such as alpha-1-antitrypsin deficiency and MAFLD.

Often described as the gold standard, primary human hepatocytes (PHHs), are sourced from donated human liver tissue, either non-transplantable donor organs or surgically resected material which can introduce substantial donor-to-donor variability and limit batch sizes. The donated tissue may be steatotic, ischemic, or diseased, all of which can compromise metabolic functionality in vitro, consequently PHHs may not fully reflect a healthy human liver population. Furthermore, any metabolic functionality remaining, for example cytochrome P450 (CYP450), is rapidly lost within a few days of culture.

As an alternative, scientists often use immortalised carcinoma cell lines (e.g. HepaRG or HepG2), which are available at scale and easy to maintain for prolonged culture times, but are prone to genetic drift and possess limited metabolic activity.

To overcome these limitations, ioHepatocytes provide a rapidly maturing, highly consistent, and scalable source of physiologically-relevant human iPSC-derived hepatocytes, powered by opti-ox deterministic cell programming. ioHepatocytes maintain metabolic function for over 17 days in 2D monolayer culture and express key Phase I, II, and III drug metabolism genes alongside functional CYP enzymes (including CYP3A and CYP2B6). When challenged with known hepatotoxins, ioHepatocytes successfully mirror the drug-induced liver injury (DILI) response of primary human hepatocytes. By combining this predictive accuracy with availability at scale, they offer an optimised cell model for hepatotoxicity screening, drug metabolism studies, and disease modelling.

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Learn more about ioHepatocytes and explore the data

ioHepatocytes display classical cobblestone morphology with prominent nuclei

ioHepatocytes rapidly acquire a homogenous hepatocyte phenotype upon thawing. This consistent and rapid maturation provides scientists with the confidence in the reproducible generation of functional cultures.

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Human iPSC-derived ioHepatocytes overcome the limitations of primary human cells and immortalised cell lines

bit.bio_ioHepatocytes_bulk-rna-seq_benchmarking_phase-I

While immortalised cell lines (e.g. HepG2 or HepaRG) display abnormal karyotypes and low metabolic activity, primary human hepatocytes (PHHs) suffer from donor-to-donor variability and lose functionality within days in 2D cultures; ioHepatocytes are a consistent, functional source of human iPSC-derived hepatocytes stable for a longer experimental window.

Transcriptome analysis of bulk RNA-sequencing data of ioHepatocytes reveals high lot-to-lot consistency and a gene expression signature highly similar to PHHs. Unlike heterogeneous populations generated by directed differentiation protocols, ioHepatocytes express mature pan-hepatocyte markers associated with hepatocyte identity and metabolic pathways, including Albumin (ALB), Hepatocyte Nuclear Factor 4 Alpha (HNF4α), ASGR1, and SERPINA1.

By integrating physiologically relevant, human iPSC-derived ioHepatocytes into their workflows, scientists can identify drug toxicity much earlier in the preclinical pipeline. This early insight helps de-risk drug discovery programs and significantly reduces the likelihood of costly, late-stage candidate failures.

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Transcriptomic profiling of Phase II and Phase III drug metabolism enzymes

bit.bio_ioHepatocytes_bulk-rna-seq_benchmarking_phase-II+III

ioHepatocytes have a high similarity in expression profile with primary human hepatocytes. Specifically, ioHepatocytes cluster closely with PHH and HepaRG and separately from the more immature HepG2 and fetal liver tissue. Importantly, ioHepatocytes express the key enzymes CYP3A4, CYP2B6, CYP2C9, GSTA1/2 and UGT1A1, ABCB11 and ABCC2.

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Human iPSC-derived ioHepatocytes recapitulate complex human liver function and secrete albumin

bit.bio_ioHepatocytes_Alb_secretion_ELISA_day11+day17

In addition to displaying a classic cobblestone morphology, prominent nuclei, and expressing core hepatocyte markers from day 4, ioHepatocytes recapitulate key functionality of mature human hepatocytes.

As the cells mature in culture, they exhibit increasing metabolic competency, demonstrated by robust and sustained albumin secretion. By successfully mirroring human biological responses, such as lipid accumulation and storage of glycogen ioHepatocytes provide a human, physiologically-relevant tool for modelling complex metabolic syndromes, such as metabolic dysfunction-associated steatotic liver disease (MASLD), and metabolic profiling.

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ioHepatocytes have functional CYP450 enzymes and can be used to model drug-induced liver toxicity in vitro

ioHepatocytes_DILI_predictivity_PHH_7testcompounds

When challenged with compounds known to cause drug-induced liver injury (DILI), the ioHepatocytes demonstrate a cytotoxic response similar to primary human hepatocytes (PHHs). Data shows similar cytotoxic response in both cell types except for Atorvastatin, which shows higher cytotoxicity in ioHepatocytes. Please note that Atorvastatin is known to be cytotoxic in 3D models of hepatocytes but not 2D models (Proctor et al. 2017), suggesting ioHepatocytes are better suited to model cytotoxic response to this drug.

This validation data confirms their suitability as a predictive human model for assessing drug toxicity in vitro and de-risking compounds early in the development pipeline.

What do scientists say about ioHepatocytes

Stuart Rushworth

Scientific Group Leader, Molecular Haematology | University of East Anglia

"We used bit.bio ioHepatocytes to validate our murine models in a human cellular context, defining a metabolic pathway in cancer. The ioHepatocytes were straightforward to use and responded to external stimuli as expected."

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ioHepatocytes

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Product resources

Brochure

ioHepatocytes

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Poster

Development of human iPSC-derived hepatocytes for drug discovery, translational research and toxicity testing

Harris-Brown et al.

bit.bio

2026

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Talk

Development of human iPSC-derived hepatocytes for drug discovery, translational research and toxicity testing

Dr Gianmarco Mastrogiovanni | Principal Scientist - Cell Type Development | bit.bio

Human Cell Forum 2025
Session 2 | bit.bio insider: Tools, tips, and what’s coming next

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Frequently Asked Questions (FAQs)

What is the functional role of hepatocytes in the liver?

Hepatocytes are the primary parenchymal cells of the liver, accounting for 80% of the liver's mass. They are responsible for essential metabolic functions, including lipid and glucose metabolism, storage of macronutrients, secretion of plasma proteins, detoxification and xenobiotic metabolism.

How do human iPSC-derived hepatocytes compare with immortalised cell lines?

Immortalised cell lines such as HepG2, often derived from hepatocellular carcinomas, exhibit abnormal karyotypes and have limited metabolic activity. ioHepatocytes overcome these limitations by providing a physiologically relevant model that expresses mature markers and displays robust functional activity. Transcriptomic analysis shows that ioHepatocytes cluster more closely with primary human hepatocytes (PHHs) than with common carcinoma cell lines.

How do ioHepatocytes compare to traditional directed differentiation?

Traditional differentiation protocols from iPSCs into hepatocytes are complex and lengthy (>30 days), frequently yielding a highly heterogeneous, immature "foetal-like" cell population. Powered by opti-ox technology, ioHepatocytes are deterministically programmed, resulting in rapid maturation, defined populations, and lot-to-lot consistency.

What is the functional experimental window for ioHepatocytes?

Unlike primary human hepatocytes (PHHs) that rapidly de-differentiate and lose Cytochrome P450 activity within a few days when in 2D culture, ioHepatocytes provide consistent functionality over 17 days. This extended experimental window enables scientists to study chronic drug exposure and include later time points in toxicity and metabolic assays.

What functional metabolic assays are compatible with ioHepatocytes?

ioHepatocytes are compatible with a wide array of standard toxicological and metabolic assays. The cells have been validated for measuring robust albumin secretion and evaluating functional Cytochrome P450 enzyme activity.

References

Ewart, Lorna, et al. "Performance assessment and economic analysis of a human Liver-Chip for predictive toxicology" Communication Medicine, 154, 2022, doi.org/10.1038/s43856-022-00209-1
Soldatow, Valerie, et al. "In vitro models for liver toxicity testing." Toxicol Res (Camb), 2(1):23–39, 2012, doi: 10.1039/C2TX20051A
Moeller, Timothy, et al. "Assessment of Compound Hepatotoxicity Using Human Plateable Cryopreserved Hepatocytes in a 1536-Well-Plate Format." Assay Drug Dev Technol, 10(1):78–87, 2012, doi: 10.1089/adt.2010.0365