Workshop on In Vitro Toxicology Adopting Integrated Discrete Multiple Organ Co-culture (IdMOC)
16th-18th Feb, 2011
Organized By
Department of Endocrinology
University of Madras
Guindy Campus
Chennai – 600 025
Tamil Nadu
International Faculty Dr. Albert P. Li & Dr. Utkarsh Doshi AP Sciences, Inc., Columbia, MD 21045, USA
Sponsored By Mahatma Gandhi-Doerenkamp Center (MGDC) for Alternatives to Use of Animals in Life Science Education, Bharathidasan University, Tiruchirappalli 620 024 People for Animals, Chennai &, Doerenkamp-Zbinden Foundation, Switzerland
About MGDC Mahatma Gandhi Doerenkamp Center (MGDC) for Alternatives to use of Animals in Life Science Education, an internationally funded research-cum- educational facility established at Bharathidasan University, Tiruchirappalli, Tamil Nadu, India, and an organ of Doerenkamp-Zbinden Foundation, Switzerland, is involved in promoting the concept of alternative approaches in biomedical testing while, at the same time, propagating the pedagogy of humane teaching in life science and biomedical science education. To popularize the pedagogical idea, MGDC has chosen to conduct seminar-cum-workshops all over the sub-continent and, as of now, has conducted 18 workshops in India and one in Ethiopia. Further, to generate man-power in the field of in vitro toxicology, the Center organizes wet lab workshops, with leading experts, either at its in vitro toxicology lab or in collaboration with other facilitators of in vitro research (
www.bdu.ac.in/mgdc).
Relevance of the workshop In Vitro toxicology The use of non-animal test methods, including computer-based approaches and in vitro studies, provides important tools to enhance our understanding of hazardous effects of chemicals and for predicting these effects on humans. In vitro tools are used principally for screening purposes and for generating comprehensive toxicological profiles, to obtain mechanism-derived information, and to provide important non-invasive tools to enhance the extrapolation from in vitro to in vivo in humans. Over the last decades an increasing number of test systems for evaluating the possible toxicological hazard of chemical compounds have been developed. Many of the systems do not rely on the use of intact animals, but make use of biological systems such as organisms in a lower level of organization, isolated organs, cell cultures, and sub-cellular systems. These in vitro systems have been extremely useful in studying the molecular basis of a chemical’s biological activity, including its mechanism(s) of toxic action. Other important developments have been achieved in the prediction of biological reactivity on the basis of a compound’s physico-chemical properties, such as structure, molecular size, reactive groups, etc, using in vitro tools. One application of this knowledge is in the construction of structure–activity relationships. The increasing possibilities to use cell and tissue cultures to measure these biomarkers of effect are now becoming complemented by the potential use of information derived from micro-array analyses, genomics, transcriptomics and proteomics.
Advantages of in vitro assays using primary cells In vitro assays involving primary cells are used routinely to evaluate organ-specific toxic effects, for instance, the use of primary hepatocytes to evaluate hepatotoxicity. Technologies for culturing primary cells from human and animal organs have been developed and have long been applied toward the evaluation of drug toxicity. Human primary cells are especially important as experimental systems for the prediction of human-specific drug properties. Applications of human hepatocytes in drug development include the evaluation of metabolic stability, metabolite profiling and identification, drug–drug interaction potential, and hepatotoxic potential. The use of intact human hepatocytes, because of the complete, undisrupted metabolic pathways and cofactors, allows the development of data relevant to humans in vivo. Incorporation of key in vivo factors with the intact hepatocytes in vitro may help predict human in vivo drug properties.
In vitro toxicity assays as an approach to define human-specific xenobiotic toxicity
Human-specific xenobiotic toxicity, whether as a result of drug metabolism or target cell sensitivity, by definition, cannot be accurately evaluated with nonhuman animal models. Short of experimentation in humans in vivo, toxicity testing in vitro using experimental systems with relevant human-specific properties represents the only practical preclinical approach to derive human-specific information for the accurate prediction of human xenobiotic toxicity.
The use of dedifferentiated cell lines such as transformed or immortalized mouse or human fibroblasts may not be useful, as neither of the above critical properties is present. The use of human organ-derived primary cells as monogenic culture (single cell type cultures) will only allow the evaluation of the effect of xenobiotics on a specific cell type which may or may not possess significant human xenobiotic metabolism pathways.
One cell system that represents the target cells and has human metabolism capacity is human hepatocytes. This cell system has the following advantages:
Human xenobiotic metabolism: Fresh isolates or cryopreserved fresh isolates of human hepatocytes are known to contain most, if not all, of the in vivo hepatic xenobiotic metabolism capacity.
Human target cells: The hepatocytes are the cells in the human liver that are damaged by hepatotoxicants, leading ultimately to liver failure.
However, a commonly used cell line, the HepG2, does not have these properties and, therefore, would not represent a relevant in vitro model for the investigation of hepatotoxicity and, more importantly, is derived from a human adenocarcinoma of the liver, not the parenchymal cells which are the in vivo target of hepatotoxicants.
Integrated Discrete Multiple Organ Co-culture (IdMOC) The IdMOC is a novel technology developed in AP Sciences laboratory as an in vitro experimental system for the evaluation of human xenobiotic metabolism, distribution, and toxicity. It is based on the concept that in the human body, there are multiple organs that are physically separated but are interconnected by the systemic circulation. The systemic circulation allows multiple organ interactions. An example of multiple organ interaction is the metabolism of a toxicant by the liver, with the resulting metabolites entering the systemic circulation, leading to the exposure of distal, nonhepatic organs to these metabolites, resulting in toxicity in these distal organs.
In vitro assays contribute to use of lesser number of animals in toxicity testing Last but not the least in vitro system addresses the ethical and pedagogical issues of animal use in biomedical testing and research. Several in vitro methods have adequately replaced the classical in vivo toxicity testing. The use of cell systems to measure responses to xenobiotics has been widely investigated in the quest for alternative methods in toxicity testing and have proved to be accurate for some testing like in ecotoxicity, skin corrositivity test, ocular irritancy/corrosivity test, etc.
Program outline
Lectures and laboratory demonstrations/hands-on training for the following:
1. Cytotoxicity assays: Single cell type and IdMOC
a. Necrosis: Cellular ATP contents
b. Mitochondrial toxicity: MTT metabolism
c. Oxidative stress: GSH content
d. Apoptosis: Caspase activation
2. P450 inhibition: Microsomes
3. P450 inhibition: Hepatocytes
Registration Fee
Rs. 10,000/- for representatives from Industries/Toxicology Laboratories
Rs. 5,000/- for university & college teachers and stipendiary research scholars
Rs. 3,000/- for non-stipendiary research scholars and post-graduate students.
Travel, Food and Accommodation Food, accommodation, and local transport will be provided free-of cost. Support for travel, by train, permissible as per the rules of University of Madras, for a few deserving participants will be considered if support is forthcoming from Funding Agencies.
Enrollment will be limited Enrollment will be limited to 40 participants, selected based on the extent to which the training will be useful in research and/or career
How to apply The intending participants should pre-register in an e-mail to <j_arunakaran@rediffmail.com>, with their CV and a brief writ-up about how the training in the workshop will be useful to them in research and/or career, as attachments, on or before 31.12.2010. Those who are selected will be informed by e-mail, on or before 07.01.2011. These candidates should finally register by sending DD for the registration charge, on or before 15.01.2011.
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