Name: NEW! ioHepatocytes
Brand: 不良研究所
SKU: io1097

不良研究所

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不良研究所_ioHepatocytes_bulk-rna-seq_benchmarking_phase-II+III

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不良研究所_ioHepatocytes_RTqPCR_panel_NANOG_G6PC_ARG1_AFP_ALB

cat no | io1097 Early Access

ioHepatocytes

Human iPSC-derived hepatocytes

Cryopreserved human iPSC-derived cells powered by opti-ox, that are ready for experiments in days
Ideal for hepatotoxicity screening, drug metabolism studies, and disease modelling
Consistent, functional hepatocytes for long-term 2D culture

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A maximum number of 20 vials applies. If you would like to order more than 20 vials, please contact us at orders@不良研究所.

Human iPSC-derived hepatocytes

Powered by 辞辫迟颈-辞虫鈩� deterministic programming, ioHepatocytes deliver a scalable, highly consistent source of human iPSC-derived hepatocytes that eliminate donor variability.

ioHepatocytes display classic cobblestone morphology with distinctive nuclei and well-defined borders. They express key pan-hepatocyte markers, including ALB, HNF4A, ASGR1, CYP3A4, and present a transcriptomic signature similar to primary human hepatocytes (PHHs).

In 2D monolayer cultures, ioHepatocytes exhibit long-term functional viability exceeding 17 days, characterised by robust albumin secretion and active CYP450 enzymes.

Critically, the cells express key Phase I, II, and III drug metabolism genes. When challenged with known hepatotoxins, they successfully mirror the DILI response of primary human hepatocytes, offering an optimised, human-relevant platform for predictive toxicology, metabolic profiling, and disease modelling.

Benchtop benefits

Consistent

Generate reliable data with ioHepatocytes that offer lot-to-lot consistency to ensure reproducible screening assays.

Functional in 2D

Evaluate chronic toxicity and compound risk with primary-like hepatocyte toxicity curves and metabolic functionality sustained over 17 days.

Defined

Assess human-relevant mechanisms using defined iPSC-derived hepatocytes expressing Phase I-III drug metabolism genes

Cells arrive ready to plate

ioHepatocytes are delivered in a cryopreserved format and are programmed to mature rapidly upon revival in the recommended media. The protocol for the generation of these cells is a three-phase process: 1) Stabilisation for 1 day 2) Pre-maintenance for 9 days 3) Maintenance of hepatocytes for remainder of assay requirements.

At 不良研究所 we have validated these culture conditions up to day 17.

Product specifications

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Starting material

Human iPSC line

Karyotype

Normal (46, XY)

Seeding compatibility

6*, 12, 24*, 48, 96* & 384 well plates
*Tested by 不良研究所

Shipping info

Dry ice

Donor

Caucasian adult male, age 55-60 years old (skin fibroblast),

Vial size

>5 x 10鈦� viable cells

Quality control

Sterility, protein expression (ICC) and gene expression (RT-qPCR)

Differentiation method

opti-ox deterministic cell programming

Recommended minimum seeding density

600,000 cells/cm虏

User storage

LN2 or -150掳C

Format

Cryopreserved cells

Product use

ioCells are for research use only

Applications

Predictive hepatotoxicity
DILI screening
Drug Metabolism and Pharmacokinetics (DMPK)
Disease Modelling

Applications

Advanced In Vitro Platforms & Organ-on-a-Chip
Functional genomics & target validation

Scale your study with volume pricing

Enabling scientists to use human cells in their research, running additional experiments without rationing cells or limiting experimental scale

Order quantity	Total vials received	Pricing tier
1 - 9 packs	2 - 18 vials	Standard price
10 - 49 packs	20 - 98 vials	Automatic 10% discount
> 50 packs	> 100 vials	> Contact us for a quote

Academic pricing: Academic users can purchase any ioCells in 3-vial packs ($/鈧�/拢 999 per pack), available year-round with any cell type combination.

What scientists say about ioHepatocytes

Stuart Rushworth

Scientific Group Leader, Molecular Haematology | University of East Anglia

"We used 不良研究所 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."

Highly characterised and defined

ioHepatocytes display classical cobblestone morphology with prominent nuclei

ioHepatocytes rapidly acquire a homogeneous hepatocyte phenotype upon thawing that is maintained throughout the culture period. Brightfield images on day 4, 11 and 17.

ioHepatocytes display binucleation

Brightfield images of day 9 post-thaw ioHepatocytes exhibiting classic hepatocyte features including cobblestone morphology, well-defined cellular borders and binucleation (yellow arrows). Original magnification 10x.

Immunocytochemistry shows protein expression of key hepatocyte markers

Immunofluorescent staining on day 4, 11 and 17 post-revival demonstrates robust expression of the hepatocyte markers, albumin, HNF4A and A1AT.

RT-qPCR shows gene expression of key hepatocyte markers

Relative gene expression at days 4, 11, and 17 post-thaw confirms rapid loss of pluripotency (NANOG) and progressive maturation towards a functional hepatic phenotype. Markers for gluconeogenesis (G6PC), urea cycling (ARG1), and plasma protein synthesis (albumin) significantly increase at days 11 and 17, while the fetal marker AFP downregulates over time.

Transcriptomic profiling of Phase I cytochrome P450 enzymes

Hierarchical clustering of bulk RNA-sequencing data reveals that ioHepatocytes (ioHeps) share high expression profile similarity with primary human hepatocytes (PHH) across 4 independent donors (cultured for 3 days). ioHepatocytes cluster closely with functional PHH and HepaRG (cultured for 8 days), segregating distinctly from immature HepG2 cells, iPSCs, and external fetal liver tissue data. Notably, ioHepatocytes demonstrate robust expression of critical drug-metabolising enzymes, including CYP3A4, CYP2B6, and CYP2C9. Data generated from cells in continuous culture (non-cryopreserved).

Transcriptomic profiling of Phase II and Phase III drug metabolism components

Bulk RNA-sequencing of ioHepatocytes (ioHeps) demonstrates a mature drug-processing profile that clusters tightly with primary human hepatocytes (PHH; 4 donors). The heatmap highlights a distinct separation from immature HepG2 lines and fetal liver tissue. For essential conjugation and transport pathways, ioHepatocytes robustly express key Phase II enzymes (GSTA1/2, UGT1A1) alongside vital Phase III transporters (ABCB11, ABCC2). Data generated from cells in continuous culture (non-cryopreserved).

Functional activity over time

Quantitative analysis of albumin secretion over time

ELISA quantification of albumin secretion confirms a time-dependent increase in metabolic competency during cell maturation. Albumin production reaches >10碌g/million cells/24hr by day 11 post-thaw, and doubles to >20碌g/million cells/24hr by day 17, establishing a robust and healthy primary hepatocyte-like functional profile capable of maintaining metabolic competency over time.

Functional characterisation of CYP3A4 metabolic activity over time

Basal CYP3A4 activity in ioHepatocytes was evaluated at days 11 and 17 post-thaw using the P450-Glo luminescence assay. As the most abundant and catalytically dominant drug-metabolising enzyme in human hepatocytes, CYP3A4 serves as a critical indicator of functional hepatic maturity. The data demonstrates measurable basal activity at day 11 (~2x10⁶ RLU/million cells/24hrs), which more than doubles by day 17 (~5.4x10⁶ RLU/million cells/24hrs), confirming robust metabolic competency over time.

Predictive hepatotoxicity screening in ioHepatocytes

Dose-dependent cell viability tracking aligns tightly with primary human hepatocyte (PHH) benchmarking data and established DILI severity categories

ioHepatocytes and primary human hepatocytes (PHH) were exposed to 7 distinct test compounds across a broad concentration gradient for 24 hours. Cell viability was determined relative to vehicle control. Upon exposure, ioHepatocytes exhibit a clear, dose-dependent cytotoxicity profile that correlates strongly with PHH benchmark performance. Both models successfully stratify the compounds based on clinical DILI severity - capturing distinct cytotoxic drops for severe/moderate toxins (e.g., Sunitinib, Amiodarone, Ketoconazole) while displaying a stable viability profile for Valproic acid, used as a control compound. Data generated from cells in continuous culture (non-cryopreserved).

DILI predictivity

Dose-dependent cell viability tracking aligns tightly with primary human hepatocyte (PHH) benchmarking data and established DILI severity categories

Frequently Asked Questions (FAQs)

What are ioHepatocytes and how are they manufactured?

ioHepatocytes are functional human iPSC-derived hepatocytes generated using 辞辫迟颈-辞虫鈩� deterministic cell programming technology. Unlike traditional directed differentiation methods that rely on staggered applications of growth factors or cytokines, opti-ox precisely activates cell-fate defining transcription factors within every single stem cell. This converts entire cultures into a defined, homogeneous hepatocyte population, ensuring rapid maturation, highly scalable manufacturing, and unmatched lot-to-lot consistency.

How do ioHepatocytes compare to Primary Human Hepatocytes (PHHs) and immortalised cell lines (e.g., HepG2)?

ioHepatocytes bridge the gap between biological relevance and high-throughput scalability:

PHHs: While PHHs remain the conventional in vitro "gold standard," they suffer from high donor-to-donor variability, limited availability, and rapid loss of function in 2D culture. ioHepatocytes provide complete lot-to-lot reproducibility and a stable experimental window with no practical upper limit on scale.
Immortalised Cell Lines: Carcinoma cell lines like HepG2 are highly scalable but display abnormal karyotypes, incomplete functionality, and severely restricted enzyme activity. Bulk RNA-sequencing data demonstrates that ioHepatocytes cluster closely with functional PHHs and HepaRGs, segregating distinctly from immature HepG2 cells and fetal liver tissue.

Can ioHepatocytes be used for predictive hepatotoxicity testing and Drug-Induced Liver Injury (DILI) screening?

Characterisation testing shows that upon exposure to known hepatotoxic compounds, ioHepatocytes exhibit a clear, dose-dependent reduction in cell viability. This performance strongly correlates with baseline PHH data and correctly aligns with established clinical DILI severity categories. This makes ioHepatocytes an ideal, human-relevant screening system for predicting compound-induced liver toxicity early in drug discovery pipelines.

Do ioHepatocytes express Phase I, II, and III metabolic machinery?

Transcriptomic validation shows that ioHepatocytes actively express the core genetic markers necessary for comprehensive xenobiotic metabolism:

Phase I (Oxidation): Robust expression of vital Cytochrome P450 enzymes including CYP3A4, CYP2B6, and CYP2C9.
Phase II (Conjugation): Consistent expression of critical transferase enzymes like GSTA1/2 and UGT1A1.
Phase III (Excretion): Defined expression of essential bile acid and drug transport genes such as ABCB11 and ABCC2.

What is the user workflow timeline and how long can these cells be maintained in culture?

ioHepatocytes are delivered in a convenient, cryopreserved format and follow a simple, robust three-phase protocol that achieves a mature phenotype in just days:

Phase 1 (Day 0鈥�1): Thawing and stabilisation for 1 day utilizing a ROCK inhibitor.
Phase 2 (Days 1鈥�10): Pre-maintenance of the hepatocytes for 9 days.
Phase 3 (Days 11鈥�17+): Maintenance for the remainder of your specific assay window.

While traditional PHHs rapidly de-differentiate, ioHepatocytes are functionally validated to maintain morphological integrity (including classical cobblestone patterns and frequent binucleation) and strong marker expression (Albumin, A1AT, and HNF4A) up to Day 17 post-thaw and beyond - enabling chronic, longitudinal testing workflows.