Alternative Models

photo credits: Copyright Medical University Graz

Cell-based toxicity assays offer a wide range of possibilities to assess important hallmarks in drug discovery processes.

Genotoxicity

Assays for

• Mutagenesis
• Clastogenic and aneugenic changes
• DNA repair mechanism

Hemocompatibility

Assays for

• Hemolysis
• Plasmatic coagulation
• Platelet activation
• Complement activation

Immunotoxicity

Assays for

• Phagocyte function (NO generation, respiratory burst, chemotaxis, phagocytosis, surface markers)
• Panels of cytokines

Chronic Cytotoxicity

3D culture of cells on

• Microbeads in benchtop biorector
• Membranes at an air-liquid interface

CONTACT
Eleonore Fröhlich
Medical University of Graz
eleonore.froehlich@medunigraz.at

photo credits: Copyright Medical University Graz

Biological mutagenicity and cytotoxicity assays can assess the hazardous potential of complex mixtures or chemicals in a very timely manner. Therefore, studies with an appropriate combination of these assays can provide useful information about the toxic/mutagenic potential.

• AMES ASSAY
• UMU ASSAY
• CALAX ASSAY

AMES ASSAY
The Ames Assay is a fast and widely employed bacterial tool to screen for mutagenic effects of chemicals or extracts and can be performed with S9 (crude liver enzyme extract) for metabolic activation. It allows for the detection of base-pair substitutions and frame-shift mutations.

UMU ASSAY
The Umu Assay is a lesser known bacterial tool for assessment of mutagenic and cytotoxic potential with or without S9, which is based on the ability of mutagens to induce genes involved in bacterial mutagenesis via the SOS-repair pathway and is performed in S. typhimurium.

CALUX® ASSAY
The CALUX® (Chemical Activated Luciferase gene eXpression) Assays (BioDetection Systems bv, Netherlands) allow for sample testing in an eukaryotic system with a quantifiable response. The CALUX Assays use the human bone osteosarcoma cell line U-2 OS allowing for testing for genotoxicity (p53 CALUX), endocrine disruptors ((anti-)ERα CALUX, (anti-)AR CALUX) as well as cytotoxicity (Cytotox CALUX).

photo credits: Copyright Medical University Graz

CONTACT
Clemens Kittinger
Medical University of Graz
clemens.kittinger@medunigraz.at

photo credits: Copyright Medical University Graz

The Flexcell® FX5K™ Tension System functions as a computercontrolled bioreactor designed to subject cells cultured in vitro to cyclic or static tensile strains. This system utilizes regulated vacuum pressure to alter flexible-bottomed culture plates, resulting in a substrate elongation of up to 25%. It has the capability to program multiple changes in frequency, amplitude, and waveform within a single regimen, effectively replicating tissue strains and frequencies found in vivo in various cell types including muscle, lung, heart, blood vessels, skin, tendon, ligament, cartilage, and bone. A range of waveform options such as static, sinusoidal, heart stimulation, triangular, square, and customizable are available. The mechanical stimulation is carried out through four independent FlexLink remote compression and/or tension controllers designed for 6-well sized plates. The BioFlex® Culture Plates possess optical clarity, allowing for direct observation of cells using both inverted and upright microscopes. These plates offer a total growth surface area of 57.75 cm2 and come with a variety of available matrix surfaces including Amino, Collagen (Type I or IV), Elastin, Pronectin® (RGD), and Laminin (YIGSR). In addition, the Tissue Train® 3D culture system allows users to create 3-dimensional cell-seeded gels and apply uniaxial tensile strains to these gels.

CONTACT
Birgit Lohberger
Medical University of Graz
birgit.lohberger@medunigraz.at

photo credits: Copyright Medical University Graz

The avian chorioallantoic membrane (CAM) assay provides an alternative versatile and ethically less objectionable in vivo model for many applications:

• Angiogenesis
• Tissue/organs xenografts
• Tumor cell lines:
  • proliferation
  • apoptosis
  • invasion
  • metastasis
• Testing biomaterials
• Testing drugs

CONTACT
Nassim Ghaffari Tabrizi-Wizsy
Medical University of Graz
nassim.ghaffari@medunigraz.at

photo credits: Copyright Medical University Graz

Spatio-temporal development of pregnancy and placenta in animals differs substantially from human. Results from animal models to study transfer of substances across the placenta lack significance. The ex vivo perfusion of a single human placental lobule is the most reliable experimental model.

Technology

• Intact human tissue, different biological barriers
• To investigate vasoactive characteristics of drugs in a human vascular system
• To determine uptake, efflux and transfer of substances across distinct barriers
• To study transfer kinetics of substances and metabolites
• Easy accessible tissue, no ethical concerns exist

Charactaristics

• A reliable model for the transfer of substances from the mother to the fetus in late pregnancy
• To imitate late placental physiology ex vivo over several hours
• A dual system with completely separated maternal/fetal circuits
• Physiological conditions for different circuits: medium, plasma, constant body temperature, pressure control and variable O2 concentrations
• Placental tissue from different pathophysiologies available

CONTACT
Christian Wadsack
Medical University of Graz
christian.wadsack@medunigraz.at

photo credits: Copyright Medical University Graz

Animal-free culture models are mandatory for translational research and the clinical application of cell culture products. Consequently, human platelet lysate is replacing the previously used fetal bovine serum in various research fields, especially for mesenchymal stroma cell expansion and in other areas of regenerative medicine. The Department of Blood Group Serology & Transfusion Medicine is offering a pooled human platelet lysate for research only, manufactured from fresh platelet concentrates in ISO 9001:2015 certified laboratories. The “O platelet in AB plasma” human platelet lysate is highly standardized and aliquots are delivered with an extensive quality certificate. On special request, a GMPgrade O/AB HPL is also available.

CONTACT
Claudia Bernecker
Medical University of Graz
c.bernecker@medunigraz.at

photo credits: Copyright Medical University Graz

Dummies like the pig’s head, the silicone model of a rabbit ear or rat and mouse dummies help to pursue the targets of reducing the burden (severity level) on the animals by nonexperienced persons and the number of animals effectively used by making frequent repetitions of a technique possible. Techniques like venous blood extraction from the ear (replaceable rubber tube), placing a venous/arterial catheter in the “ear vein”/”artery of the ear”, placing an arterial catheter in the common carotid artery or endotracheal intubation can be trained on the pig head dummy. Practicing blood sampling on rabbits itself is a big burden on the animal. The silicone model of the ear is a proven replacement method to train this technique. Rat and mouse dummies (entire animal and skeleton dummies), which accurately display the anatomy of the animals can substantially reduce the number of animals used for autopsies.

CONTACT
Aida Saric
Medical University of Graz
aida.saric@medunigraz.at