BioIVT LIVERPOOL™ Cryoplateable and Cryopreserved Primary Human Hepatocytes

Hepatocytes provide a valuable alternative to in vivo testing. Since these cells mimic the properties of the liver, researchers can evaluate the metabolism, drug-drug interaction and toxicity of drug candidates in a cost-effective manner.

We partnered with BioIVT to provide hepatocytes which can be used in cross comparisons of metabolism and in preclinical programs.

BioIVT is a worldwide provider of biological products to life sciences and pharmaceutical companies. They specialize in control and disease state matrices manufactured from human and animal blood, plasma, serum, tissues and other fluids which are used in drug discovery, compound development, clinical and research diagnostics.

Their hepatocytes and subcellular fraction products are prepared from fresh whole tissues, no resect tissues. Products are manufactured to ensure consistent and reproducible results. All products are characterized and data is available here.

Human Hepatocyte Products

  • LIVERPOOL™ Cryoplateable Human Hepatocytes
  • LIVERPOOL™ Cryopreserved Human Hepatocytes
  • Cryoplateable Human Hepatocytes
  • Cryopreserved Human Hepatocytes

LIVERPOOL™ Cryopreserved and Cryoplateable Pooled Human Hepatocytes

LIVERPOOL™ is a patented product that is blended to ensure sufficient activity levels in all CYP categories. LIVERPOOL™ cryoplateable and cryopreserved hepatocytes are used to  conduct toxicity, inhibition, and metabolism studies. They  have high levels of induction following treatment with  known inducers.

  • Purpose-Pooled: A proprietary algorithm, drawing up a cell database of known substrate performances, selects cryoplateable hepatocytes lots from a well-characterized cell bank to achieve targeted enzyme levels.*
  • Characterization: All LIVERPOOL™ products are fully characterized across numerous substrates to ensure consistency and reliability.
  • Format Choice: Available in 5, 10, 20, 50, and 200 donor pool sizes.
  • Savings: In both time and money, LIVERPOOL™ hepatocytes enable researchers to eliminate pooling procedures, characterization testing, and costly multi-donor work.
  • Large Lot Sizes: All contributory hepatocyte lots are sourced from whole, intact, transplant-quality livers, ensuring each donor lot can create large product sizes. This enables researchers to have great consistency across multiple experiments.
  • Quality: All LIVERPOOL™ lots must meet and exceed stringent quality control specifications prior to release.
  • Reproducibility: Allows multiple experiments at different times from a single donor.

* The process for producing the LIVERPOOL™ pooled human hepatocyte products is covered by one or more U.S. or foreign patents and patent applications, including U.S. Patent No. 7,604,929.


20-Donor LiverPool

Targeted and actual data enzyme characterization from a LiverPool 20-donor purpose-pooled cryopreserved hepatocyte lot.

induction with prototypical inducers

Cryoplateable Primary Human Hepatocytes

Cryoplateable human hepatocytes are available from male and female donors. Hepatocytes sourced from BioIVT are isolated from fresh, whole livers and cryopreserved the same day. Cryoplateable human hepatocytes are used for long-term studies like biliary excretion, induction and toxicity. More recently, scientists also have been using cryoplateable hepatocytes to study transporter uptake, viral transduction and gene induction. BioIVT cryoplateable human hepatocytes have high levels of induction following treatment with known inducers omeprazole (CYP1A2), phenobarbital (CYP2B6), and rifampin (CYP3A4). All plateable lots have greater than 80% viability and 70% confluency on day 5.

  • Convenience – No waiting for fresh livers
  • High Quality – Established responsiveness to inducers
  • Full Characterization – Known phenotypes and donor demographics
  • Reproducibility – Allows multiple experiments at different times from a single donor
  • Easy-to-Use – Allows multiple donor studies in the same experiment
  • High Throughput – Use in 96-, 384- and 1536-well plates

Induction with Prototypical Inducers as Measured by Enzyme Activity
Induction with Prototypical Inducers as Measured by Enzyme Activity

Cryopreserved Primary Human Hepatocytes

With activity equivalent to that of freshly isolated hepatocytes, cryopreserved hepatocytes from BioIVT let you conduct your experiments whenever you want. Hepatocytes are isolated and cryopreserved on the same day. Cryopreserved hepatocytes typically are used to study both phase I and phase II metabolism.

Human cryopreserved hepatocytes are genotyped for the following: NAT1, NAT2, CYP2C9, CYP2C19, and CYP2D6. Donor demographics and medical histories are provided.

  • Convenience – No waiting for fresh tissue
  • Testing – Characterized for a wide range of enzyme activities
  • Reproducibility – Long shelf life allows multiple experiments at different times from a single donor
  • Ready-to-Use – Thaw and use when you’re ready
  • Quality – Cells have >80% viability

Click here to get a complete list of currently available hepatocyte lots from BioIVT

BioIVT recommends the use of:

  • INVITROGRO™ CP medium (Cat. No. Z99029) and TORPEDO™ Antibiotic Mix (Cat. No. Z99000) for thawing and plating the cryoplateable human hepatocytes.
  • INVITROGRO™ HT medium (Cat. No. Z99019) and TORPEDO™ Antibiotic Mix (Cat. No. Z99000) for thawing cryopreserved hepatocytes.
  • INVITROGRO™ CP medium (Cat. No. Z99029) and TORPEDO™ Antibiotic Mix (Cat. No. Z99000) for thawing and plating the LiverPool™ cryoplateable human hepatocytes.
  • INVITROGRO™ HT medium (Cat. No. Z99019) and TORPEDO™ Antibiotic Mix (Cat. No. Z99000) for thawing the LiverPool™ cryopreserved human hepatocytes.

Techniques and Best Practices

The following are some key best practices in ADME-Tox in vitro techniques that BioIVT has gleaned from being a pioneer and market leader in this field since 1990.

Tip 1: Follow the correct protocol
- Different manufacturers = different protocols
- Protocols are optimized for product characteristics
The correct protocol will give better results Use INVITROGRO™ hepatocyte medium

Tip 2: Store and thaw the cells under appropriate conditions
- Store cells with liquid nitrogen at < -150°C in either liquid or vapor phase of liquid nitrogen
- Transport cells under dry ice for less than 30 minutes
- Thaw cells promptly upon removal from the storage unit
- Thaw cells in media pre-warmed to 37°C
Improper storage or thawing temperature degrades cell viability and performance

Tip 3: Minimize assay preparation
- Choose cell lots with high viability by checking the manufacturer’s tested viability or by performing the assay yourself
- BioIVT cells are guaranteed at 5 million viable cells/vial and >70% viability without Percoll® purification
- Cryoplateable Hepatocytes do not require a centrifugation and washing step prior to plating
Fewer steps means less time is required for cryoplateable hepatocytes to run your assays

Tip 4: Do not allow the cells to settle
- Hepatocytes are heavy and settle rapidly
- Invert gently for consistent aliquots

Homogenous aliquots result in consistent cell counts and seeding
consistent cell counts and seeding

Tip 5: Handle hepatocytes gently
- Hepatocytes are fragile
- Avoid rapid pipetting or trituration of the cells
Improper handling can affect cell viability and performance

Tip 6: Measure volumes precisely
- Accuracy is important when filling hemacytometer chambers for cell counting
- Add exactly 10 μL into each chamber
Varying volumes can result in more than 15% error in cell counts and can impact your results

Tip 7: Count the correct cells in the appropriate manner
- Additional cell types may be present
- Do not count the smaller non-parenchymal cells
- Only count 2 of the 4 sides within the square
Errors in cell counts can impact your results

Tip 8: Use the correct Trypan Blue concentration
- Use the recommended 1:5 dilution of Trypan Blue
Trypan Blue is toxic to cells at higher concentrations and will affect viability

Tip 9: Count the cells quickly
- After the Trypan Blue is added, count the cells within 1-2 minutes
Trypan Blue is toxic to cells longer exposure times

Tip 10: Distribute the cells evenly
- As a final step to plating, gently shake the plate in a north-south and then east-west direction to distribute the cells evenly.
Even distribution of cells results in better attachment

Tip 11: Remove loose cells
- When washing up, tap the plate lightly to help suspend the loose cells
Unattached cells may affect plated cells and reduce their adherence

Tip 12: Add media carefully to plated cell
- Monolayers of plated hepatocytes are fragile
- Carefully pipette media down the sides of the wells
Rough handling will disrupt the hepatocytes monolayer and affect experimental outcomes

remove loose cells

Tip 13: Avoid pooling issues
- When pooling hepatocytes from multiple lots, work quickly and efficiently
- Avoid delays in thawing, mixing, counting, and plating
- To avoid these problems and reduce waste, consider pre-pooled products BioIVT’s Liverpool™ Pooled
Cyropreserved Human Hepatocytes

Delays in thawing, mixing, counting, and plating can affect cell viability

Tip 14: Ask for help when needed
- Contact our Technical Support
- Request an in-house product demonstration or seminar

BioIVT Medium and Medium Supplements

Advantages and Benefits:

  • Optimized for culturing and/or plating primary human hepatocytes
  • Reliable and consistent performance across cell lines
  • Easy media preparation – just thaw, mix and use
  • Superior performance and consistent results in assays for drug metabolism, transporter activity, hepatotoxicity and P450 enzyme induction

Cryoplateable BioIVT
Cryopreserved BioIVT PHH
Pooled BioIVT PHH

Click here to get a complete list of currently available hepatocyte lots from BioIVT.

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