Thursday, 23 July 2015

Hepatocyte Solutions

Primary hepatocytes are widely used as the gold-standard in-vitro liver model for biological, pharmacological and toxicological studies in drug discovery and development.

BiopredicInternational provides a wide range of high quality fresh or cryopreserved hepatocytes, obtained from humans and animals. These hepatocytes meet the specific needs of our customers many in-vitro applications.





Key advantages of hepatocytes produced at Biopredic International include:

- A regular supply of fresh human and animal primary hepatocytes, i.e. 2 isolations per week from multiple species (rat, mouse, dog and monkey)

- A wide range of both fresh and cryopreserved hepatocytes, offering flexible choices
- Larger batch sizes of cryopreserved hepatocytes allowing scale up of experiments (i.e. pooled donors)

- Pre-qualified cryopreserved human and animal hepatocytes allowing in-vitro protocols to meet your specific application requirements

- Full characterisation data for each product lot, including Phase I /II drug-metabolizing enzyme activities 

- Professional storage and delivery of fresh plated hepatocytes with Biopredic’s patent-pending Sealovac™ Packaging Technology (guaranteed for a maximum of 3 days)

- A complete Certificate of Analysis for all products, including donor information, quality control values and specifications, as well as serological and sterility results

- Custom preparations according to your specifications.

Caltag Medsystems are the exclusive UK and Ireland distributor for Biopredic products. For further information about hepatocytes or any of their other products please call +44 (0)1280 827460 or email office@caltagmedsystems.co.uk.


Hepatocytes for Cellular Biology: A Cost Effective Alternative

Biopredic are pleased to launch a cost effective format of hepatocytes to be grown in monolayers for cellular biology. These hepatocytes are perfect for customers who wish to carry out the following applications:

- Clearance
- Metabolism
- Uptake studies
- Induction
- Cytotoxicity
- Apoptosis
- Geno toxicity

These cost effective hepatocytes are isolated from the residues of fresh hepatocyte production and are negative for HBV, HCV and HIV1/2. They are isolated using a two-step collagenase treatment. Their QC procedure includes:

- Plateability
- Viability: >70% post-thaw viability WITHOUT Percoll purification step
- Number of viable cells / vial: ≥ 5 x 10^6

Product Code: HEP187-P

Caltag Medsystems are the exclusive UK and Ireland distributor for Biopredic products. For further information about hepatocytes or any of their other products please call +44 (0)1280 827460 or email office@caltagmedsystems.co.uk.

Hepatocyte Cell Line: HepaRG

BioPredic have an exclusive license to manufacture the hepatocyte cell line: HepaRG.

HepaRG is a unique and well established hepatic cell system able to produce early hepatic progenitor cells as well as completely mature human hepatocytes.  It is an alternative to fresh hepatocytesm providing a one-stop hepatic in-vitro model for:

In-vitro ADME applications:

Inhibition assays
Drug metabolism and clearance
Metabolite ID

In-vitro toxicity testing applications:
Hepatotoxicity (reactive metabolites, lipidosis, choleastasis etc)

Virology (HBV, HCV…)
Infection with hepatitis and other viruses for replication


Innovative Modes of Culture
BioArtificial Livers 
Transgenic liver humanized animals with HepaRG™ cells
…. an ever-expanding range of applications!

HepaRG™ Key Advantages

- The capacity to produce early hepatic progenitor cells as well as completely mature human hepatocytes.
- A morphology and metabolic capacity similar to human primary hepatocytes.
- The first human hepatoma cell line to express main CYPs, functional nuclear receptors and major sinusoidal and canalicular hepatic drug transporters.
- Overcomes limited availability of liver tissue.
- Excellent batch-batch reproducibility and sub-cultures.
- CYP activities and expression of canalicular drug transporters can be increased by classical inducers.
- A robust cell-based system to evaluate metabolism of test compounds.
- A suitable model for high throughput screening of chemicals.
- A unique and exclusive model for in vitro pharmacological and toxicological studies.
- Usable and stable for at least 4 weeks once differentiation is achieved, ideal for long-term studies.
- Can support the complete replicative cycle of HBV.


In-vitro ADME Applications

Drug metabolism and clearance
HepaRG™ cells be classified as an “average human hepatocyte population” and express multiple functional Phase 1 and 2 drug metabolising enzyme (DME) activities comparable to levels in cultures of primary human hepatocytes. This makes them a much better model for metabolism and clearance studies than any other cell line currently available. The prediction of in-vivo intrinsic clearance of a panel of drugs has been shown to be mostly within 2fold using HepaRG™ cells. Moreover, the sustained presence of DMEs over weeks makes them ideal for measuring the clearance of low clearance compounds.

Metabolite ID
HepaRG™ cells have a full complement of DMEs, cofactors and transporter proteins and the profile of DME activities and expression provides a good representation of that in primary human hepatocytes. Together with the high stability of the enzymes, this makes these cells ideal for producing relevant drug metabolites in sufficient quantities to enable metabolite identification.

DME induction assays
HepaRG™ cells express functional transcription factors involved in the regulation of DMEs. These cells are responsive to prototypical inducers of CYP1A1/2, CYP2B6, CYP2C8, CYP2C9, CYP2C19, and CYP3A4 and can be used to predict in-vivo CYP3A4 induction of test compounds by constructing a calibration curve using known potent and mild inducers of this CYP (“AUC/F2” or “RIS”). The protocol for CYP induction using HepaRG™ cells has been accepted by ESAC and an OECD guideline for use of HepaRG™ cells in induction studies is in preparation.

DME inhibition assays
The high levels of DMEs in HepaRG™ cells makes them ideal for inhibition screening studies. The IC50 values of a panel of CYP inhibitors using HepaRG™ cells correlated very well with those determined in primary human hepatocytes. Time-dependent inhibition can also be investigated using these cells.

Functional transporter assays
HepaRG™ cells express a number of uptake and efflux drug transporters and form tight junctions and bile canaliculi, making them ideal for uptake and biliary secretion studies.

In-vitro toxicity testing applications

Hepatotoxicity Screening and Mechanistic Testing Applications

Carbohydrate and lipid metabolism
HepaRG™ cells display glucose and insulin responsiveness and can be used to investigate carbohydrate homeostasis and lipogenesis.

Reactive metabolites
The stable and high levels of DMEs in HepaRG™ cells make them ideal for long-term and repeated dose toxicity studies in which a compound may be bioactivated and/or cause latent effects over days or weeks.

Steatosis
The accumulation triglycerides in HepaRG™ cells is predictive of the in-vivo steatosis reported in patients chronically treated with steatogenic drugs. Lipid accumulation directly correlates with levels of mRNA encoding for lipid synthesis markers, thus this toxicity can be monitored visually and at the gene level.

Phospholipidosis
HepaRG™ cells can be used to discriminate between drugs which cause steatosis (after 24 h) and phospholipidosis (evident after 2 weeks) e.g. amiodarone; and those which cause steatosis only e.g. tetracycline.

Cholestasis
Cholestatic drugs (via direct inhibition or down-regulation) can be identified by measuring their effects on bile acid efflux in HepaRG™ cells.

Genotoxicity and carcinogenicity
Since HepaRG™ cells have a good metabolic capacity and proliferative capacity, they can be implemented into the micronucleus test (MNT), as recommended by ECVAM and IWGT workshops. Whole genome expression analysis of HepaRG™ cells can discriminate between genotoxic and non-genotoxic compounds and provide information on compound- and time-dependent effects on cell cycle and apoptosis signalling pathways.

Mitochondrial toxicology
Apoptosis
Inflammation

Virology and Infection

HBV, HCV, HEV
HepaRG™ cells support viral replication can be used to model viral infection and provide insight into the processes involved in virus receptor binding, uptake, and membrane fusion

Malaria

Innovative Modes of Culture

BioArtificial Livers (BALs)
HepaRG™ cells can be cultured in 3D bioreactors to generate an in-vivo-like assembly of highly polarised cells with high and sustained CYP activities. This makes them a promising cell type for use as liver assist devices or BALs.

Transgenic liver humanized animals with HepaRG™ cells
HepaRG™ cells can be transplanted into chimeric mice to repopulate their livers and express human primary hepatocyte–specific genes and proteins. Such a HepaRG™-mouse combination represents a promising model for in-vivo studies of drug metabolism and hepatitis virus infections.

Spheroids
Micropatterns
Dynamic fluidic models
Co-culture models
Stem cell cultures



Caltag Medsystems are the exclusive UK and Ireland distributor for Biopredic products. For further information about HepaRG or any of their products please call +44 (0)1280 827460 or email office@caltagmedsystems.co.uk

Cryopreserved Hepatocytes

Biopredic International has more than 20 years of in-house experience in cryopreserving human and animal hepatocytes.

Cryopreserved hepatocytes are offered in two formats. These are suitable for either suspension applications or plateable applications. 

Cryopreserved Human Hepatocytes for Suspension Applications

All suspension qualified human cryopreserved hepatocytes have post-thaw viabilities over 85%.  They are also characterized for P450 and Phase II drug-metabolizing enzyme activity.  Both single donor and pooled human cryopreserved hepatocytes are suitable for use in a variety of applications including metabolic stability, clearance, metabolite ID, metabolite profiling, species comparison studies, and uptake transporter studies.

Single Donor Cryopreserved Human Hepatocytes, Suspension Qualified are produced from a single human donor.  Each lot number represents a specific individual.  Demographic information can be used as a factor in the selection of specific lots of these hepatocytes.

Pooled Cryopreserved Human Hepatocytes, Suspension Qualified are a source of pooled hepatocytes, and are produced from multiple, single donors to represent an average population phenotype.  Note that pooled human cryopreserved hepatocytes should be thawed with pooled human cryopreserved hepatocyte medium for the optimum results.

Please contact Caltag Medsystems to request the current inventory or for a quotation. Email office@caltagmedsystems.co.uk or call +44 (0)1280 827460.

Cryopreserved Human Hepatocytes for Plated Applications

Plateable hepatocytes are offered in different grades according to the application required
- Cryoplateable Human Hepatocytes (Grade P)
- Cryoplateable and inducible Human Hepatocytes (Grade I)
- Cryoplateable & sandwich culture hepatocytes

All plateable cryopreserved human hepatocytes have post-thaw viabilities over 85%.  Each lot can also be plated in a collagen-coated plate or similar.  These lots are guaranteed to maintain a healthy monolayer for a minimum of 3, 5, or 7 days in culture, depending on their qualification.  The monolayer confluency must be over 85% for the duration of the culture to be qualified as a plateable lot.

Cryoplateable Human Hepatocytes (Grade P)

Cryopreserved Human Hepatocytes, Plateable, Metabolism Qualified, can be used for multiple applications, including the study of low-clearance compound metabolism.  Each lot is tested for metabolic activity of known low-clearance substrates for CYP2C9, CYP2D6, and CYP3A4.  These lots are guaranteed plateable for a minimum of 3 days in culture.

Cryoplateable and inducible Human Hepatocytes (Grade I)

Cryopreserved Human Hepatocytes, Plateable, Induction Qualified, can be used for a variety of applications; including to determine if compounds of interest are inducers of CYP1A2, CYP2B6, and/or CYP3A4.  Qualification includes testing induction of each isoform with a known inducer and measuring the fold change in mRNA expression and activity.  These lots are guaranteed plateable for a minimum of 5 days in culture.

Cryoplateable & sandwich culture hepatocytes

Cryopreserved Human Hepatocytes, Plateable, Transporter Certified can be used for multiple applications including uptake and efflux transporter assays such as facilitated active uptake and B-Clear.  These lots are characterised and demonstrate transporter uptake and efflux activity similar to fresh human hepatocytes.  Qualyst Transporter Solutions (Durham, NC) characterises these cells as experts in the hepatic transporter field.  These lots are guaranteed plateable for a minimum of 7 days in culture.

Please contact Caltag Medsystems to request the current inventory or for a quotation. Email office@caltagmedsystems.co.uk or call +44 (0)1280 827460.

Cryopreserved Animal Hepatocytes

Triangle Research Laboratories provides cryopreserved hepatocytes from a variety of clinical species, including, but not limited to:

- Cryopreserved Mouse – 24 hours
- Rat (Sprague-Dawley, Wistar Han, Wistar)
- Mouse (CD-1 and C57Bl/6)
- Dog (Beagle)
- Monkey (Cynomolgus)

Every lot of animal cryopreserved hepatocytes is characterised for viability/stability, attachment efficiency, monolayer confluency, and general Phase I and Phase II drug metabolizing enzyme activities.  There is a wide selection of lots that can be used for a variety of applications, including, but not limited to:

- Drug metabolizing enzyme activity
- Suspension drug transporter uptake
- Time-dependent metabolism (plated)
- CYP450 Induction
- Plated drug transporter assays
- Toxicity assays

Cryopreserved Animal Hepatocytes for Suspension Applications

All suspension qualified animal cryopreserved hepatocytes have post-thaw viabilities over 80%.  These lots are also characterized for general Phase I and Phase II drug-metabolizing enzyme activity.

Please contact Caltag Medsystems to request the current inventory or for a quotation. Email office@caltagmedsystems.co.uk or call +44 (0)1280 827460.

Cryopreserved Animal Hepatocytes for Plated Applications

All plateable cryopreserved animal hepatocytes have post-thaw viabilities over 80%.  Each lot can also be plated in a collagen-coated plate or similar.  These lots are guaranteed to maintain a healthy monolayer for a minimum of 3 days in culture.  The monolayer confluency must be over 85% for the duration of the culture to be qualified as a plateable lot.  These lots are also characterized for general Phase I and Phase II drug-metabolizing enzyme activity.

Please contact Caltag Medsystems to request the current inventory or for a quotation. Email office@caltagmedsystems.co.uk or call +44 (0)1280 827460.

For instructions on how to handle cryopreserved hepatocytes, including thawing and plating:
Thawing and Plating Cryopreserved Hepatocytes (2303)

For instructions to count hepatocytes with the Trypan Blue Exclusion Method:
Cell Counting with Trypan Blue Protocol (6666)


Caltag Medsystems are the exclusive UK and Ireland distributor for Biopredic products. For further information about hepatocytes or any of their other products please call +44 (0)1280 827460 or email office@caltagmedsystems.co.uk.

Fresh Hepatocytes

Freshly isolated hepatocytes are widely accepted as a superior in-vitro model for evaluating the in-vitro hepatic metabolism and toxicity of test compounds because they more closely reflect the in-vivo situation, when compared with other models.

Biopredic International has a well-organised supply network which guarantees the regular delivery of human liver resects and animal livers. Human liver is obtained from partial hepatectomies and from whole livers rejected for transplantation. High quality fresh human and animal hepatocytes are isolated from the livers and then fully characterized.

Fresh human hepatocytes products include:

- Hepatocyte suspensions suitable for short-term metabolism studies (up to 4 hours)

- Plated (monolayer) hepatocytes cultures for longer-term metabolism studies (up to 3 days).

Hepatocytes suspensions suitable for short-term metabolism studies (up to 4 hours)

In order to provide suspended human hepatocytes, freshly isolated hepatocytes from human liver tissue are rapidly suspended and preserved in Biopredic’s optimized hypothermic medium to maintain cell viability and functional activity during shipment  (up to 48hours).

Key features of human fresh hepatocytes in suspension

- Good availability of fresh cells with proprietary production process
- High quality product combined with optimized hypothermic shipping condition
- Stable and retained viability and expression level of metabolizing enzymes 
- Greater flexibility to handle cells either in suspension or plated cell culture
- Characterization data available for each product batch sheet 
- General patient information (age, gender, race, health conditions, serology)
- Characterization for fresh human hepatocytes in suspension

Biopredic provides the following data for each lot of hepatocyte suspension:

- Viability after isolation and after cold shipping period, yield, plateability after cold storage period, morphology before and after shipment
- Phase I drug-metabolizing enzymes with a cocktail of probe substrates: CYP1A2/ 3A4/ 2B6/ 2D6/ 2C19
- Phase II drug-metabolizing enzymes: UGT1A1/ UGT1A6/ UGT1A9/ UGT2B15/ SULT1A1/ SULT1A3/ 4/ SULT1E1
- Serological and sterility control data

Shipment of fresh human hepatocytes in suspension

Freshly isolated hepatocytes are suspended in an optimized hypothermic medium and then aliquoted into cryovials containing at least 10 million cells per vial (1ml).The cell suspension is shipped the same day of the preparation (or the following day) at hypothermic condition (2-8°C).

The viability, plateability and functionality of preserved cell suspension are well maintained for at least 48h at 4°C.

Fresh Hepatocytes Email Alert Service

To receive dates of intended surgeries and to reserve hepatocytes, please sign up here.

To view the available fresh suspension hepatocytes products, please click here.

Plated (monolayer) hepatocytes cultures for longer-term metabolism studies (up to 3 days)

To provide adherent monolayer cultures, freshly isolated and viable cells are plated at a desired seeding density onto collagen-coated culture supports according to the customer’s request/ordering.

After overnight attachment, the seeding medium is replaced with either a short-term culture medium or long-term culture medium (Lanford et al., 1989, journal and vol, page?) to produce short-term (≤ 3 days) or long-term hepatocytes (≤ 6 days) cultures in which cells retain a differentiated phenotype for at least 1 week.

Both freshly prepared short-term and long-term cultured hepatocytes contain all major phase I and phase II enzymes as well as the whole gene regulation pathway necessary to investigate the potentiality of drugs to induce phase I and phase II enzymes.

Key features of human fresh plated hepatocytes

- Multi-format plated cells and custom preparation available
- Greater viability and confluence of cells in monolayer
- High expression levels of major hepatic drug-metabolizing enzymes, co-factors and cell membrane receptors
- Greater flexibility for applications with either short or long-term cultures
- Optimized packaging and shipping conditions with Biopredic's patented technology - SEALOVAC
- Full characterization data available for each product batch sheet
- General patient information (age, gender, race, health conditions, serology)
- Characterization of human fresh plated hepatocytes

Biopredic International provides the following data for each lot of plated hepatocytes:

- Cell viability and cell culture period after isolation and seeding
- Cell morphology and culture confluence by microphotography
- Phase I drug-metabolizing enzymes with a cocktail of probe substrates: CYP1A2/ 3A4/ 2B6/ 2D6/ 2C19)
- Phase II drug-metabolizing enzymes: UGT1A1/ UGT1A6/  UGT1A9/ UGT2B15/ SULT1A1/ SULT1A3/ 4/ SULT1E1
- Serological and sterility control data

Shipment of human fresh plated hepatocytes

Plated hepatocytes cultured into multi-well plates are firstly filled with either short-term or long-term culture medium (with or without Phenol red) according to customer’s specification/request.

The pre-filled plate is then packaged using Biopredic's patent-pending Sealovac™ Technology for the shipment at room temperature (18-30°C).
Cell quality and functional activity of packaged monolayer cells are well maintained for at least 48hours.

Fresh Hepatocytes Email Alert Service

To receive dates of intended surgeries and to reserve hepatocytes, please sign up here.

To view the available fresh monolayer hepatocytes products, please click here.

Fresh animal hepatocytes

Biopredic International provides a wide range of fresh animal hepatocytes obtained from a variety of species: rat, mouse, hamster, rabbit, cynomolgus monkey, dog, minipig and other species (available on demand).

Fresh animal hepatocytes can be prepared from either single animal liver donor (such as mouse, rat, dog, monkey, rabbit or hamster) or pooled from several animal donors (mouse, rat or rabbit) according to our production schedule.

All rodent hepatocytes are isolated from animals housed in our animal facility which is managed in strict compliance with the Directive 2010/63/EU of the European Parliament and of the Council.

Hepatocytes of other animal species are isolated from the liver tissues collected following approved protocols and respecting all ethical and legal requirements.

Fresh animal hepatocytes products include:
- Hepatocytes suspensions
To view the available fresh suspension hepatocytes products, please click here.

- Plated (monolayer) hepatocytes cultures
To view the available fresh monolayer hepatocytes products, please click here.

Fresh Animal Hepatocytes Email Alert Service

To receive alerts when fresh animal hepatocytes are available, please sign up here.


Screen CETP inhibitors and measure CETP activity

Cholesteryl Ester Transfer Protein (CETP) transfers cholesteryl ester from HDL to LDL or VLDL in exchange for triglyceride, therefore CETP inhibition is viewed as an attractive approach to elevate HDL.

BioVision's CETP Inhibitor Screening Kit II provides a fluorometric assay for the screening, studying, and characterisation of mammalian CETP inhibitors.

The kit contains enriched human CETP, instead of the rabbit serum controls supplied in most kits. The controlled and consistent level of CETP allows for greater reproducibility and increased sensitivity compared to using rabbit serum, which may suffer from variability and low CETP concentration.

For user convenience, the kit also includes the inhibitor control- Anacetrapib, which is not provided in any other kit.

Features & Benefits:

  • The most advanced kit on the market: includes human CETP enzyme for high reproducibility and increased sensitivity
  • CETP inhibitor Anacetrapib included for ultimate user-convenience
  • Fluorometric assay provides enhanced sensitivity and specificity
  • Simple assay procedure
  • Adaptable to High-Throughput assays
  • Suitable for multiple sample types, including: cell and tissue culture supernatants, urine, plasma, and serum.
Product code: K594-100

BioVision also provide the CETP Activity Fluorometric Assay Kit II for measuring CETP Activity in animal serum, plasma or recombinant protein.

Features and Benefits:
  • Simple, rapid & convenient assay
  • Torcetrapid is included as an inhibitor control to validate the assay
  • Positive control (rabbit serum) also included
Product code: K595-100

The CETP Activity and Inhibitor kits use a self-quenched fluorescent neutral lipid that can be measured when transferred to an acceptor molecule. The fluorometric intensity is directly proportional to the amount of neutral lipid transferred.

Caltag Medsystems is the UK distributor for BioVision. If you have any questions about these products please email office@caltagmedsystems.co.uk, or click here to view Biovision's full product range.

Friday, 17 July 2015

Silensomes: One single model for DMPK phenotyping studies

Silensomes™ are human pooled liver microsomes in which one specific CYP has been chemically knocked out and are capable of directly quantifying and predicting the contribution of CYP enzymes in drug metabolism.

Until now, a battery of in-vitro tests recommended by the regulatory agencies (EMA/FDA) are required for this CYP phenotyping assay. These tests (correlation analysis, antibody or chemical inhibition and metabolism by recombinant human enzymes) have a number of disadvantages:

- No direct quantitative measurement of the contribution of each CYP in the metabolism of a drug (correlation analysis)
- Model is not fully representative of the liver enzyme profile (e.g. human recombinant CYP450)
- Lack of specificity (e.g. anti-CYP antibodies and chemical inhibitors)

To overcome the disadvantages of the current methodologies, Biopredic International has developed a patented new in-vitro drug development model.

Silensomes™ are validated human pooled liver microsomes (HLMs) chemically and irreversibly inactivated for one specific CYP using mechanism based inhibitors (MBI).

Each Silensomes™ is available as ready-to-use HLMs chemically knocked-out for one specific CYP activity (1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1, 3A4) with each showing high specificity and efficiency of their targeted CYP inhibition (>80%), and only minor impact (<20%),

Key Advantages of Silensomes™:
- Silensomes™: the model of choice for determination of the CYP contribution all along the development plan of a new chemical entity

- Irreversibly inhibited CYPs

- For initial rate conditions

- For saturating conditions

- One single model for CYP phenotyping assays, instead of a battery of in-vitro tests

- Quantitative: the true contribution of the CYP450 in the metabolism of the coumpound can be measured

- A powerful model with an excellent specificity, potency, stability and predictability to ensure reliable extrapolation of the drug-drug interaction risk

- More representative of in-vivo, using human liver microsomes rather than recombinant enzymes isolated from non-mammalian cells

Silensomes™ are available for compound screening purposes and also for regulatory validation

Silensomes are now available for beta testing. Please contact us if you would be interested in this.

Caltag Medsystems are the exclusive UK distributor of Biopredic products. Please call the office on +44 (0)1280 827460 for further information about their products, or email office@caltagmedsystems.co.uk.

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