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PRIMACYT - a large Portfolio of Transporter Assays,
ADME and Toxicological Assays
for fast and cost-effective Screening.


Start your Project and why you should contact us.
Primacyt is a functional precision service organization in the field of OATPs. We offer a broad portfolio of transporter assays for OATPs. We support industry, research organizations and government in the field of in vitro studies related to drug interaction from the beginning of your projects until preclinical completion.

We are highly committed to offering additional categories of laboratory services and analytical topics as a commitment to the continuous expansion of our leading position in the industry. We are your partner and the partner of your clients in the education, training, and development of testing protocols. We also offer permanent routine lab services as a customized approach and service. You are in a position to open up additional revenue streams for your organization and partners.

Our partners contact us for getting advice on many questions and facts related to

  • Drugs that cause liver damage,liver damage from medication, drug induced liver injury,
  • toxic liver, liver toxicity symptoms, side effects of liver damage
  • hepatotoxin, drug induced hepatitis, hepatotoxicity symptoms, hepatic toxicity,signs of hepatotoxicity, analysis of hepatotoxicity, or hepatotoxic drugs.


We are your partner when it comes to questions about drug-drug interaction or related lab service work for drug development and more.
Hepatoxicity in 30 seconds.
The liver is known as one of the most important sources of metabolism and drug biotransformation. It is logical that liver cells are the choice for toxicological and pharmacological testing.

The liver is fundamental for the transformation and clearance of chemicals and is responsive to the toxicity from such compounds. All these chemicals are called hepatotoxins. Overdoses and even lower rates than recommended may cause liver injuries. Chemical agents, natural chemicals such as microcystins or herbal remedies can cause hepatotoxicity. Chemicals that cause liver injury are called hepatotoxins.

Specific medicinal agents, when taken in overdoses and sometimes even when introduced within therapeutic ranges, may injure the organ. Other chemical agents and solvents, such as those used in laboratories and industries, natural chemicals (e.g., microcystins) and herbal supplements can also induce hepatotoxicity.
Nonsteroidal anti-inflammatory drugs, antiviral drugs for HIV infection, compounds used in chemotherapy

Symptoms of hepatotoxicity are manifold and comprise nausea, vomiting, abdominal pain, loss of appetite, diarrhea, atypical tiredness, weakness, yellowing of the skin and eyes (jaundice), and abnormal swelling or weight gain exceeding about 2 – 3 kg per week.

The diagnosis of liver damage requires a series of options:
Blood tests, examination of the level of bilirubin in the blood and extracellular,
Testing the level of elevated liver enzymes,

Treating liver damage means in the first instance to stop medications which are processed by the liver. An alternate choice is prescribing medications for reducing the symptoms such as swelling by increasing urinate fluid.

The availability of high throughput in vitro liver models would be a great resource and enable the shift to greater use of the alternative toxicity testing methods. In this review, we discuss the benefits and pitfalls of traditional liver-derived in vitro systems and examine novel in vitro liver models and their usefulness for toxicity testing.

A summary of commonly used in vitro hepatotoxicity model systems and novel in vitro hepatotoxicity model systems can be found at the following link below:


Further details about In Vitro Platforms for Evaluating Liver Toxicity are available here, describing classifications of drug-Induced human liver injury, assays and markers reported in primary literature to address physiological mechanisms of toxicity in multi-component liver platforms that include human hepatocytes, and examples of clinical compounds selected for validating liver platform toxicity responses.

For more details, please click below:

Project Schedule - How we can work together
Primacyt Cell Culture GmbH provides cell culture solutions for biomedical and cell biology research. PRIMACYT is focused on in-vitro-technologies, certified according to GLP. Our focus is in the fields of hepatocytes, subcellular fractions, skin tissues, cell culture media, hydrogel 3D cultures, and collagen products. We provide solutions for human and animal research.

According to the needs of your organization, we can agree on a mutual confidentiality agreement. We discuss your project needs and send you the first offer with the contractually relevant project details.

Our past cooperation partners foresee different needs for the formal contracts the parties need to agree upon. As typically our joint projects are relatively uncomplicated relative to drug discovery projects, we suggest a simple, fast-track approach to save your time for the benefit of your projects.

We are in a position to provide umbrella or master agreement type contract versions, followed by a project amendment or packaging everything in a single document. We are prepared to send you templates for such agreements including material transfer agreements if needed. Contract languages can be English or German for you as a commercial or government / academic entity.

We always suggest to exchange data through cloud mechanisms and to cooperate by life video or telephone conferences also in other languages including Chinese.


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In Vitro Model Systems
Summary of classically in vitro hepatotoxicity model systems.

The list below contains five different model with different throughput rates and certain important other attributes.

Model
Model Throughput Consideration
Other
Performance
Liver slices
Fairly high
Maintenance of liver structure, zone-specific CYP activity, toxicity mechanisms
Good in in vitro / in vivo correlation of xenobiotic metabolism
Immortal Hepatic Cell Lines
Application dependent
Lack of phenotypic and functional characteristics of tissue
Retains expression of many liver-specific functions
Primary Hepatocyte Suspensions
Fairly high
Better estimate of internal clearance, but loss of certain cell properties
Retains high level of enzyme functionality (close to in vivo)
Primary Hepatocyte Cultures
Application dependent
Induction/inhibition of the metabolizing enzymes can be studied. Cells are capable of re-establishing cell-cell interactions and polarity
Inability to maintain in vivo liver-specific functionality for long-term culture

Primary Hepatocyte Cultures – Sandwich
Application dependent
Better maintenance of liver-specific functionality
• Prevents loss of viability
• Functional bile canaliculi

Loss of liver-specific functionality, morphology and phenotype in long-term cultures

Novel in-vitro Model Systems
Summary of novel in vitro hepatotoxicity model systems.

The list below contains five different model with different throughput rates and certain

Model
Model Throughput Consideration
Other
Performance
Multi-well perfused Bioreactor
High
Uses greater cell numbers and larger media volumes,
has been validated with rat and human hepatocytes.
Cells form 3D tissue constructs,
sustained liver-like cell,
good correlation with in vivo clearance rates.
Bioartificial Liver
Low
Requires large number of cells, limited cell types.
Allows for studies of functional heterogeneity,
ability to evaluate hepatotoxicity using human blood.

Primary Cell Co-cultures
Application dependent
Best results shown with non-hepatic (e.g., fibroblasts) cells. Methods vary greatly among laboratories.
Improved hepatocyte function,
allows for studies of immune-mediated toxicity. Good correlation with in vivo toxicity.
Embryonic Stem Cells
Application dependent
Ethical concerns. Low expression levels of liver-specific metabolism genes.
Defined phenotype
Induced Pluripotent Stem Cells
Application dependent
Complex reprogramming steps
• Low expression levels of liver-specific metabolism genes
Defined phenotype
• Allows studies of inter-individual variability.
Hepatoxicity Assays
Overview

Various liver-derived in vitro model systems are available for the investigation of potential adverse effects of chemicals and drugs.

Primary hepatocytes, liver tissue slices, isolated microsomes, perfused liver, and immortalized cell lines are in use. Primary isolated liver cells and immortalized cell lines are currently most widely used for testing liver toxicity.

Limited throughput, loss of viability, and decreases in liver-specific functionality and gene expression are common shortcomings of these models.

Alternate solutions are three-dimensional tissue constructs, bioartificial livers as well as embryonic- and adult tissue-derived differentiation of stem cells into hepatic lineage-like cells. The latter approach offers the open supply of hepatocytes from multiple individuals helping to improve reproducibility and enabling the testing of individual-specific toxicity.

Co-cultures of various cell types in combination with hepatocytes aim to preserve liver-specific morphology and functionality beyond those provided by cultures of pure parenchymal cells.
Typical Assay Applications

Assesment of compounds

Overview

For hepatotoxicity testing, hepatotoxicity/cytotoxicity assays are suited to measure cell death due to the cytotoxic effect of small molecules compositions for all kinds of applications including pharmaceuticals, industrial chemicals, and consumer products.
Assays utilize Primary human hepatocytes and HepaRG cells as viable surrogates and are typical for these types of studies.

Hepatotoxicity in vitro safety testing in early stages of drug discovery

Toxicity assays in vitro simulate in vivo tissue studies and are a reliable tool for safety evaluation in early stages of drug discovery. Preferentially, to address the need for early stage hepatotoxicity testing, single and multiplexed hepatotoxicity assays represent effective in vitro models to assess hepatotoxicity potential in cells derived directly from human liver tissue.
Switching to studies in vivo is a reasonable option at the late stage of development when the number of suited compounds has been limited.

High Content Analysis (HCA) assays

Image-based HCA assays are based on cryopreserved primary human hepatocytes or HepG2 (human liver hepatocellular carcinoma) cells. These analyses provide a thorough assessment of hepatotoxicity while supplying detailed mechanistic information underlying the observed toxicity.
Additional assays are available for drug-induced cholestasis, steatosis, or mitochondrial toxicity.

Predictive in-vivo Assays
Overview

Predictive in vitro assays for early toxicity evaluation are extremely important for improving the drug development process. Reducing drug attrition rates during clinical development is mandatory.

In vitro toxicity assays based on induced pluripotent stem cell (iPSC)-derived hepatocyte are useful as efficient tools for safety and efficacy testing for the improvement of drug development efficiency. Such models offer the benefit of having a primary tissue-like phenotype, sufficient availability, and the potential to compare cells from different individuals.

Predictive assays are powerful when high throughput screening is possible and multiple parameters can be derived of general and mechanism-specific hepatotoxicity.

Parameters of interest can be such as cell viability, nuclear shape, average and integrated cell area, mitochondrial membrane potential, phospholipid accumulation, cytoskeleton integrity, and apoptosis.

The approach allows the processing of a large number of compounds, and the determination of mechanisms of toxicity, facilitating the safety assessment of drugs and chemicals.

In vitro Hepatotoxicity Services

As the primary organ involved in drug detoxification, consequently the suffers most from drug toxicity. Drug-induced liver injury (DILI) is the leading cause of drug failures.

Hepatotoxicity implies chemical-driven liver damage. It can be induced by a drug itself or indirectly by the generation of reactive metabolites.

Sometimes during biotransformation, some of the drug or xenobiotic may be activated to chemically reactive species, i.e. reactive metabolites. This biotransformation of relatively inert chemicals to highly reactive intermediary metabolites is commonly referred to as metabolic activation or bioactivation, and it is known to be the initial event in many chemically induced toxicities.

Reactive metabolites undergo oricesses of redox cycling and oxidative stress /ROS generation or lipid peroxidation yielding GSH depletion. These acticities and the alteration of Ca2+ homeostasis lead to metabolic alterations yielding atopsosis and necrosis. Covalent binding triggers timmune response including the inactivation of biomolecules.

Summarizing the effects, toxic injury to hepatocytes is produced through multiple mechanisms involving damage to biomolecules, alteration of cell homeostasis/function and cell death.



2D-based Hepatotoxicity Assay

Toxic injury to hepatocytes is produced through multiple mechanisms involving damage to biomolecules, alteration of cell homeostasis/function and cell death.

Especially the 2D-cultures of hepatocytes, offer advantages relative to human hepatocytes as gold standard.

Advantages allow savings in costs and time preferably at high throughput / low compound rates and provide the generation of more parameters related to toxicity generation and its mechanisms. Resulting assays can be combined with test articles to analyze cytotoxic effects and multiple dilutions of the full dose including benefits from modified concentration ranges for the determination of the optimal range.

Possible assay parameters include the evaluation of cell viability, apoptosis, cellular stress, cytoplasmic membrane disruption and mitochondrial membrane potential.



3D-based Hepatotoxicity Assay

3D-based assays use spheroids and are exceeding the benefit of 2D-based assays for drug safety testing.

Long-term 3D cell cultures deliver improved analysis of the complex in vivo microenvironment. Long-lasting toxicity studies for several are possible, resulting in a higher level of reliable data regarding drug safety.

The hepatocyte spheroids are exposed to compounds to be tested and can include positive and negative controls for a predefined length of time.

The availability of multi-parametric data, extended to ATP content and other endpoints, can be monitored and analyzed to improve the quality of data and the prediction of drug hepatotoxicity. Combined readouts can better rule out false negative candidates who might not exhibit toxic effects with a single parameter.

It is possible to derive multiple data from 3D cultures pending on the number and size of spheroids formed. Glutathione, ATP content, mitochondrial membrane potential, and oxidative stress may be of relevance.



High Throughput / High Content Screening

High-throughput toxicity screening

With the advantage of simultaneously screening multiple candidate compounds, High-throughput toxicity screening is ideal for the dose-dependent bioactivity evaluation of chemicals in a cost and time efficient way.

High-content toxicity screening
Since drug toxicity is usually a combined phenotype of multiple mechanisms, single experimental approaches are not capable of capturing the complexity involved in cellular toxicity.
Cell-based high-content cytotoxicity screening that can provide multi-parametric information on cellular toxicity using automated fluorescence imaging.


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