Replace Animals in Australian Testing

Alternative Methods

In Vitro

 

Alternative to Fetal Calf Serum (FCS)

VitroGro (animal product free, growth-enhancing factor technology) can be used instead of FCS. It enhances “…cell replication, protein synthesis and migration” http://www.tissuetherapies.com/

Zet (Centre for Alternative and Complementary Methods to Animal Testing ) have compiled a catalogue of commercially available formulations, structured for their specified fields of application, i. e. cells/tissues/cell lines and manufacturers. Serum Free Cell Culture Media - Updated Product Guide 1/2004-05 provides an overview of the range of commercially available serum-free media for cell culture.
http://www.zet.or.at/subnode,6,294,de,Serum_Free_Cell_Culture_Media,publikationen.php 

For an overview of serum-free media, see http://www.focusonalternatives.org.uk/PDFs/serum_free_cell_culture.pdf

 

Drug screening

Synthetic and natural products are evaluated in vitro as potential anticancer agents and to combat the AIDS virus. In particular, the U.S. National Cancer Institute (NCI) has developed a Human Tumor Cell Line Screen and an AIDS Antiviral Screen.
http://nci.chemfinder.com/

 

Toxicology

In vitro methods can be used to identify potential toxic hazards in the following areas:

•       Biological products and vaccines

•       Cardiac toxicity

•       Embryotoxicity

•       Endocrine disruptor

•       Eye and Skin Irritancy

•       Genotoxicity

•       Liver and intestine toxicity

•       Pyrogenicity (fever producing)

Biological products and vaccines

Cell and tissue cultures are used to test a number of pharmaceutical products, including vaccines, antibiotics, therapeutic proteins, toxins and antitoxins. (Eg. SNAP-25/Endopeptidase assay for Botulinum Toxin Potency Testing)
http://www.nc3rs.org.uk/downloaddoc.asp?id=853&page=1023&skin=0

Cardiac Toxicity

Early assessment of the cardiac safety of compounds can be done using the in vitro hERG electrophysiology assay.
http://www.avivabio.com/cardiac-safety.php

InVitroHeart is a project using human embryonic stem cells. The aim is to create pre-validation in vitro models and reduce the use of animal experimentation for cardiotoxicity testing.
http://www.invitroheart.org/

Embryotoxicity

ECVAM Scientific Advisory Committee (ESAC) has endorsed the following methods:

  • Embryonic stem cell test for embryotoxicity
  • Micromass embryotoxicity assay
  • Whole rat embryo embryotoxicity assay

Although these cannot replace the animal tests, they could contribute to reducing animal use. (ESAC Statements, May 1, 2002)
http://www.alttox.org/ttrc/toxicity-tests/repro-dev-tox/

Endocrine Disruptor

In vitro tests can detect for chemicals that act as estrogen agonists or antagonists.
(eg. Certi-Chem ER Assay, LUMI-CELL® ER Assay)

Eye and Skin Irritancy

Agarose Diffusion Method tests for toxicity of plastic and synthetic devices are used in medical devices such as heart valves, artificial joins and intravenous lines, using human cells.

Agarose Diffusion Method can also be used to test for eye irritancy.

Cottine, m. et al. 1993, Critical evaluation of an agarose diffusion method for the assessment of eye irritancy. ATLA 21, pp 427-440
http://altweb.jhsph.edu/publications/journals/atla/21_4/atla21_4b.htm

Synthetic tissues and human skin recombinants provide a non-animal alternative for testing skin corrosion and toxicology. (Eg. Corrositex®, EpiDerm™, EpiOcular™, Episkin™, TestSkin® etc)

Genotoxicity

The Ames test (a bacterial test for mutagentic properties of chemicals) is the most widely used.

( Manahan , S. E., 2002, Toxicological chemistry and biochemistry. CRC Press, p.175. )

There are a number of in vitro tests modeled on the Ames test which use eukaryotic cells and human cells and are thus more accurate. For eg. GreenScreen™ 

http://www.gentronix.co.uk/Products/GreenScreenHC/tabid/68/Default.aspx

Olaharski, A., et al., 2009, Evaluation of the GreenScreen GADD45α-GFP indicator assay with non-proprietary and proprietary compounds. Mutation Research/Genetic Toxicology and Environmental Mutagenesis. 672(1), pp 10-16.

The in vitro micronucleus test could be used as an alternative to the chromosome aberration test or the mouse lymphoma assay in the regulatory genotoxicity battery.

Lorge, E. et al., 2007, Genetic toxicity assessment: employing the best science for human safety evaluation. Part II: Performances of the in vitro micronucleus test compared to the mouse lymphoma assay and the in vitro chromosome aberration assay. Toxicol Sci., 96(2), pp 214-217
http://toxsci.oxfordjournals.org/cgi/reprint/kfl193v1

Liver and Intestine Toxicity

LIINTOP (Liver/Intestine Optimisation) is a project that aims to evaluate and improve in vitro models for pharmacokinetics and pharmacodynamics.
http://www.liintop.cnr.it/index.php

VitroCellomics is another project which uses human embryonic stem cells. The aim is to develop in vitro models that reliably reflect human hepatic properties, thus reducing the use of animal experiments for predicting drug metabolism and toxicity.
http://www.vitrocellomics.org/

Pyrogenicity

Interagency Coordinating Committee on the Validation of Alternative Methods (ICCVAM) reviewed five in vitro tests for detecting the pyrogenicity of pharmaceuticals and other products. They found that although none of these can completely replace the rabbit pyrogen test (RPT), they can reduce the number of animals used in RPT.
http://iccvam.niehs.nih.gov/methods/pyrogen/pyrogen.htm

 

Some Australian suppliers of In-Vitro Products:

Auspep Pty Ltd
1800 805 393
www.auspep.com.au
auspep@auspep.com.au

Agenix Ltd
08 9478 4753
www.agenix.com

BioCore Pty Ltd
1800 003 991
www.biocore.com.au
sales@biocore.com.au

Bioscientific Pty Ltd
1300 246 724
www.biosci.com.au

CSL Limited (which has incorporated Zenyth Therapeutics)
1800 063 892
www.csl.com.au
customerservice@csl.com.au

Dermatest Pty Ltd
02 9597 7115
www.dermatest.com.au

John Morris Scientific
www.johnmorris.com.au

In Vitro Technologies Pty Ltd
1300 552 003
www.invitro.com.au
care@invitro.com.au

Invitrogen Australia Pty Ltd
1800 331 627
www.invitrogen.com
mail.australasia@invitrogen.com

Millenium Science
1800 678 242
www.mscience.com.au
customerservice@mscience.com.au

Serotec
03 9662 7348
www.csiro.au/places/ps122.html
George.Lovrecz@csiro.au

 

The Human Tissue List February 2009 includes 267 records of various tissues available in Australia   Web link

Australian Human Tissue Banks

For Brain research:

For Cancer research:

See also: 'Tumour banks: providing human tissue for cancer research' The Medical Journal of Australia, 175, 2001, pp.293-4. 

For Heart research:

•       St Vincent’s Institute of Medical Research and The Victor Chang Cardiac Research Institute Myocardial Tissue Bank

For Leukaemia and Lymphoma:

•       The PricewaterhouseCoopers (PwC) Foundation Leukaemia and Lymphoma Tissue Bank
http://www.leukaemia.org.au/web/research/tissuebank.php

For M.S.:

•       University of Sydney, Multiple Sclerosis Brain Bank
http://www.msbrainbank.org.au/

For bone and tissue grafts:

  • Australian Biotechnologies

http://www.ausbiotech.com.au

  • Perth Bone and Tissue Bank Inc

http://www.perthbonebank.com

For disease pathogenesis:

  • Fetal tissues from Diabetes Transplant Unit, Prince of Wales Hospital, Sydney,

Professor Bernard E Tuch: b.tuch@unsw.edu.au

For alcohol research:

  • Human 'Brain Bank', Tissue Resource Centre, Neuropathology Unit of the Department of Pathology, University of Sydney

http://www.perthbonebank.com

Australian Bioengineering of Tissues

For liver transplants:

Chihara, Y et al, 'Primary human hepatocytes on biodegradable poly (l-lactic acid) matrices: A promising model for improving transplantation of tissue engineering' Liver Transplantation, 17/2 Feb 2011 (pp104-114).

For breast reconstruction after breast surgery:

Australian Tissue Engineering Centre

http://www.atecltd.com.au

For kidney diseases:

Song, B et al, 'Generation of induced pluripotent stem cells from human kidney mesangial cells', Journal of the American Society of Nephrology, 22/7, 2011, pp.1213-20. 

For disorders of the endometrium:

Ye, L et al, 'Generation of human female reproductive tract ep0ithelium from human embryonic stem cells', PLoS one, 6/6. 2011, e21136

Also see 3-D bio-printing: the human tissue printer by Invetech

http://www.invetech.com.au

    

 

Useful Links

CarcinoGENOMICS In vitro methods for assessng the carcinogenic potential of compounds

http://www.carcinogenomics.eu

 

ESTIV European Society of Toxicology in Vitro promotes in vitro toxicology in scientific studies and education in Europe.

 

IIVS Institute for In Vitro Sciences is dedicated to the advancement of alternative methods worldwide

http://www.iivs.org

 

IVTS In Vitro Toxicology Society is a discussion forum for scientists on in vitro toxicology.

http://www.estiv.org

 

Sens-it-iv provide in vitro alternatives to animal tests currently used for the risk assessment of potential skin or lung sensitisers

http://www.sens-it-iv.eu

Last reviewed: 25 October, 2011