Cell Products ⇢ Wild-type cells ⇢

Ulti-HEP: Wild-type iPSC-derived human hepatocytes

Accurately model liver biology in vitro with functionally mature iPSC-derived human hepatocytes that maintain similar physiological behavior as primary human hepatocytes.

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Revolutionizing Liver Models for Drug Discovery

The pharmaceutical industry urgently needs efficient, human-relevant in vitro liver models that can seamlessly integrate into drug discovery pipelines.

These models are crucial for accurately identifying potential drug candidates and evaluating their toxicity profiles.

Traditional liver models, such as cancer cell lines or primary cells, fall short due to inherent limitations in functionality, scalability, and consistency. This is why we have created Ulti-HEP, human induced pluripotent stem cell (iPSCs) derived hepatocytes, which overcomes the limitations of traditional models.

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Why Ulti-HEP?

Our cells are extensively characterized to meet the demanding criteria of the pharmaceutical industry, Ulti-HEP provides a robust platform for advancing liver research, ensuring better predictions, and accelerating the journey from discovery to treatment.

Human-relevant functionality

Highly metabolic, replicating the natural processes of human hepatocytes which last much longer than primary cell lines in culture.

High performance

Are well suited for predictive toxicology and drug efficacy screening assays. Unlike primary cells, Ulti-HEP can also be genetically edited in order to create bespoke disease models too.

Consistent & scalable

Designed to deliver reliable, reproducible results for research since they are sourced from a single donor. Researchers can select from several donors to suit their research needs.

Ulti-HEP’s Advantages

   Derived from induced pluripotent stem cells (iPSCs), our Ulti-HEP cells exhibit all human-specific metabolic pathways, giving you relevant and reproducible data, leveraging the consistency of iPSC culture.

   Express comparable levels of liver maturity markers to primary human hepatocytes

   Express higher levels of urea cycle markers and secrete higher levels of urea compared to liver carcinoma cell lines

  Ulti-HEP demonstrate a functional gluconeogenesis pathway

   Demonstrate comparable levels of CYP450 markers and CYP3A4 activity to primary human hepatocytes

   They show functional localization and function of ASGR1 for GalNAc-dependent drug deliveries

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Technical Data

Morphology

Ulti-HEP demonstrate the characteristic hepatocyte cobblestone morphology.

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Figure 1: Representative cell morphology pictures of induced pluripotent stem cells (iPSCs), hepatocellular carcinoma HepG2 cells, DefiniGEN Ulti-HEP, and primary human hepatocytes (PHH). The pictures reveal the characteristic cobblestone morphology of Ulti-HEP, and the presence of a uniform monolayer following >3 weeks of iPSC differentiation. Objective: 20x.

Maturity marker analysis

Ulti-HEP express similar levels of liver maturity markers compared to primary human hepatocytes (PHH).

Hepatocyte maturity markers - ICC photos v2
Figure 2: A) Representative immunocytochemistry pictures and protein quantification showing expression levels of the hepatocyte maturity markers albumin (red), alpha-1-antitrypsin (A1AT; green), HNF4A (green), and AFP (red) in liver carcinoma HepG2 cells, Ulti-HEP, and primary human hepatocytes (PHH; 3 donors). Cells were counterstained with DAPI, and data are presented as mean ±SEM of n=3-4 independent experiments.
Hepatocyte maturity markers - ICC
Figure 2: B) Representative immunocytochemistry pictures and protein quantification showing expression levels of the hepatocyte maturity markers albumin (red), alpha-1-antitrypsin (A1AT; green), HNF4A (green), and AFP (red) in liver carcinoma HepG2 cells, Ulti-HEP, and primary human hepatocytes (PHH; 3 donors). Cells were counterstained with DAPI, and data are presented as mean ±SEM of n=3-4 independent experiments.

Urea cycle marker analysis

Ulti-HEP express higher levels of urea cycle markers and secrete higher levels of urea compared to liver carcinoma cell lines.

Urea cycles enzymes - WB photos v2

Figure 3: A) Protein expression levels of the urea cycle enzymes OTC, ASS1, ASL, CPS1, and ARG1 in liver carcinoma HepG2 cells and Ulti-HEP. 

Urea secretion
Figure 3: B) Protein expression levels of the urea cycle enzymes OTC, ASS1, ASL, CPS1, and ARG1 in liver carcinoma HepG2 cells and Ulti-HEP. B) Urea secretion in liver carcinoma HepG2 cells and DefiniGEN Ulti-HEP. Data are presented as mean ±SEM of n=3-4 independent experiments.

Functional gluconeogenesis

Ulti-HEP demonstrate a functional gluconeogenesis pathway and respond to gluconeogenesis inducers.

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Figure 4: A) Simplified schematic of the gluconeogenesis pathway within human liver.

G6PC basal mRNA levels - HepG2_Ulti-HEP_PHH
Figure 4: B) G6PC mRNA levels in liver carcinoma HepG2 cells, Ulti-HEP, and primary human hepatocytes (PHH). 
G6PC induction - mRNA levels
Figure 4: C) G6PC mRNA levels in liver carcinoma HepG2 cells and Ulti-HEP treated with 0.1mM dbcAMP (gluconeogenesis inducer).
HepG2 vs Ulti-HEP - Glucose secretion
Figure 4: D) Glucose secretion in dbcAMP-treated liver carcinoma HepG2 cells and Ulti-HEP upon pyruvate challenge. Data are presented as mean±SEM of n=3-4 independent experiments. mRNA expression data were normalized to 18S rRNA.

CYP450 expression and activity

Ulti-HEP demonstrate comparable levels of CYP450 markers and CYP3A4 activity to primary human hepatocytes.

CYP450 mRNA levels - Basal expression

Figure 5: A) mRNA expression levels of Phase I CYP450 genes in liver carcinoma HepG2 cells, Ulti-HEP, and primary human hepatocytes (PHH).

Basal CYP3A4 activity

Figure 5: B) Basal CYP3A4 activity in liver carcinoma HepG2 cells, Ulti-HEP, and PHH. mRNA data were normalized to the housekeeping gene 18S rRNA and are presented as mean ±SEM of n=3-4 independent experiments. CYP3A4 activity data were normalized to ATP levels and are presented as mean ±SEM of n=3-5 independent experiments. For PHH data, cells from 3 independent donors were used.

ASGR1 expression and function

Ulti-HEP demonstrate functional membrane localization and activity of the Asialoglycoprotein receptor 1 (ASGR1).

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Figure 6: A) Representative immunocytochemistry pictures showing the localization of ASGR1 in the Ulti-HEP membrane. Cells were counterstained with the membrane marker E-cadherin and DAPI.

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Figure 6: B) The effect of ASGR1 in the transport of GalNAc-siRNA conjugate targeting GAPDH in Ulti-HEP using GalNAc-Cy3 staining and qPCR. Data are presented as mean±SEM of n=3-4 independent experiments. mRNA expression data were normalized to 18S rRNA. NTC: non-template control.

Key Publications

Our Ulti-HEP have been used in several peer-reviewed publications, you can learn more about how they have helped to accelerate research below:

Scientific paper

Modeling Disease with Human Inducible Pluripotent Stem Cells

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Scientific paper

Production of hepatocyte-like cells from human pluripotent stem cells

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Scientific paper

Generation of Hepatocytes from Pluripotent Stem Cells for Drug Screening and Developmental Modeling

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Frequently asked questions

Do Ulti-HEPs express ASGR1?

Yes, ASGR1 and ASGR2 has been detected by qPCR analysis and ICC and has been shown to localize on the cell membrane. 

Do they have a functional urea cycle?

Yes this has been characterized by gene expression profiling, Western blot analysis and Ornithine stimulated functional analysis.

Can you see CYP expression activity?

Expression of CYPs is detectable by qPCR analysis, but they are lower than Primary Human Hepatocytes (PHH).