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Accepting the challenge to advance more humane science
 
 

PAST IFER RESEARCH GRANT RECIPIENTS

Since its founding in 1985, IFER has supported projects for the development, validation and implementation of scientifically valid methodologies that advance scientific research, product testing and education without harming animals. IFER has fulfilled this mission by making shrewd investments in better science.

IFER’s ventures have been made in neither stocks nor bonds or commodities and futures; the investment “risks” made by IFER have involved cutting edge research methodologies and technology at the early stages of their development. In addition to providing grants to principle investigators, IFER has invested in scientists at the earliest stages of their careers through the Graduate Fellowship Program started in 1996. IFER has also supported efforts to disseminate information designed to increase awareness and proficiency in the use of alternatives through conferences, publications and workshops.

The following pages reflect Principal Investigators and their studies, which IFER has supported.


1987-1988 Awards

Tchao, Ruy, Ph.D.,
Philadelphia College of Pharmacy and Science.
Development of a Trans-epithelial Permeability Assay as an Alternative to the Draize Eye Test

We have investigated several types of detergents. The effects of these detergents on the integrity of the epithelium closely match their ranking of irritancy in the rabbit eye test as published in the literature. In other words, the measurement of permeability of a sheet of epithelium produced in vitro by MDCK cells could be used as an assay for the eye irritancy of certain chemicals.

Because this new method measures the functional activity of an epithelium instead of cytotoxicity, we have been able to rank several mild detergents which have eluded other in vitro test systems using cytotoxicity as a measurement.

Other advantages of this method are that it is simple, rapid, quantitative, and reproducible. It requires no animals and would be more economical than the Draize rabbit eye test.

 

Babich, Harvey, Ph.D. and Borenfreund, Ellen, Ph.D.,
The Rockefeller University.
Use of Metabolising Systems in the Neutral Red Assay to Develop an Alternative to the Draize Rabbit Skin Tolerance Test

We have investigated the use of the human liver cell line, HepG2, as both the chemical activator and target cell in the Neutral Red Assay. The HepG2 cell line was selected as it retains some of the drug and chemical metabolizing capabilities of normal liver cells. The ability to perform an in vitro cytotoxicity assay with a cell line of human origin would make such an assay more relevant and pertinent for assessment of chemical risk to human health.

 

1988-1989

Tchao, Ruy, Ph.D.,
College of Pharmacy and Science, Philadelphia, PA.
Validation of the Trans-epithelial Permeability Assay as an Alternative to the Draize Eye Test

The subject of this year’s proposal is validation. A large selection of commercial products, including shampoos, fragrances, and cosmetic products will be used.

 

Isseroff, Roslyn Rivkah, Ph.D.,
University of California, Davis.
Alternative Method to Whole Animals for the Evaluation of Commercial Products: Monitoring the Production of Leukotrienes by Cultured Human Keratinocytes in Response to Toxic/Irritant Subjects

This project is an in vitro test designed to replace animals and human volunteers in testing skin irritants. The approach is to evaluate the potential for cutaneous irritation of commercial products via a cell culture system. Human keratinocyte cultures, initiated from neonatal foreskins discarded at routine circumcision, will be incubated with known strong cutaneous irritants. The generation of radiolabeled leukotrienes by the cells after exposure to these irritants will be monitored utilizing both reverse and normal phase HPLC methodologies.

 

Babich, Harvey, Ph.D. and Borenfreund, Ellen, Ph.D.,
The Rockefeller University, New York, NY.
Develop an in vitro test that duplicates the metabolizing capacity of intact skin together with the Neutral Red Assay

The research project will use cultured human skin keratinocytes and human skin fibroblasts, singly and in combinations, in the in vitro neutral red cytotoxicity assay for assessing the dermatotoxicities of chemical test agents. The in vitro model will assess cellular damage to skin (i.e., the “corrosive” response) rather than the immune inflammatory response. An important aspect of this experimental design is the use of cell types of human origin, rather than from experimental laboratory animals. Thus eliminating the need for extrapolation from animals to humans.

 

1989-1990

Benson, G. John, Ph.D.; Wilson, W.A., Ph.D.; and Thurmon, J.C., Ph.D.,
University of Illinois, Urbana, IL.
Develop computerized case simulations for teaching anesthetic case management

 

Prokop, Ales, Ph.D.,
Washington University, St. Louis, MO.
Development of Dynamic, Tissue-like Density Hepatocyte Culture as an Alternative to Animals for Toxicity Evaluation

 

Babich, Harvey, Ph.D. and Borenfreund, Ellen, Ph.D.,
The Rockefeller University, New York, NY.
Use of Metabolizing Systems in the Neutral Red Assay to Develop an Alternative to the Draize Rabbit Skin Tolerance Test

 

Phelps, John, Ph.D.,
University of Texas Health Science Center at Houston.
Assessment of the effectiveness of an interactive video laser disc as a replacement of a live animal physiology lab on cardiac output

 

Van Buskirk, Robert, Ph.D.,
Department of Biological Sciences, State University of New York, Binghamton, NY.
Multiflour: A Multipotential In Vitro Approach to Cytotoxicity

 

1990-1991

Van Buskirk, Robert, Ph.D.,
Department of Biological Sciences, State University of New York, Binghamton, NY.
Develop a multi-endpoint assay for a human skin equivalent (year 2)

 

Lucas, Jen Hill, Ph.D.,
Center for Network Neuroscience, Department of Biological Sciences, University of North Texas, Denton.
A Prototype Rapid Intervention Protocol for the Treatment of Acute Phase Physical Injury to the Spinal Cord

 

Koushanpour, Esmail, Ph.D.,
Northwestern University, Chicago, IL.
An Interactive Computer Simulation Program of Renal Function

 

1991-1992

Cohen, Perrin, Ph.D. and Block, Martin, Ph.D.,
Northeastern University, Boston, MA.
Validation of non-animal methods in undergraduate psychology courses

 

Van Buskirk, Robert, Ph.D.,
Department of Biological Sciences, State University of New York, Binghamton, NY.
To conclude development of the human skin equivalent for multiple endpoint assays project. This will include cryopreservation of synthetic human skin cells

 

Haschek-Hock, Wanda M., Ph.D.,
University of Illinois, Urbana, IL.
Develop a lung culture system to replace the whole animal for evaluation of antifibrogenic agents. This will assist in testing air born toxicants

 

Smeak, Daniel D., Ph.D.,
The Ohio State University, Columbus, OH.
Evaluation of a hollow organ simulator as an alternative in teaching veterinary students

 

1993-1994

Van Buskirk, Robert, Ph.D.,
Department of Biological Sciences, State University of New York, Binghamton, NY.
Evaluation of Human Corneal Models for Automated In Vitro Toxicology

 

Stoll, Lynn, Ph.D.,
University of Iowa
Use of Coculture Systems as Cell Culture Models for Study of Vascular and Respiratory Function

 

Bauer, Michael S., Ph.D., and Seim III, Howard B., Ph.D.,
Colorado State University, Fort Collins, CO.
Development and validation of an electric psychomotor skills board as an objective evaluation tool and model for teaching basic psychomotor and surgical skills

 

1994-1995

Van Buskirk, Robert, Ph.D.,
Department of Biological Sciences, State University of New York, Binghamton, NY.
Development of an artificial liver model for automated in vitro toxicology

 

McGovern, Kathy Ann, Ph.D.
Department of Radiation Oncology, Division of Cancer Biology, University of Arizona Health Sciences Center
Development of versatile tissue model. Characterization and evaluation of a cell culture system for toxicity testing by nuclear magnetic resonance spectroscopy

Objectives: “ (1) to develop a hollow fiber reactor for controlled growth of cells in three dimensions in imitation of a perfused tissue, (2) the further development of a magnetic resonance method for monitoring oxygen concentration within this reactor, and (3) to optimize/characterize conditions for growth.” During the first phase of this project, these objectives were accomplished, and a tissue model in which cells can grow in three dimensions surrounding an artificial capillary bed was developed. A variety of magnetic resonance methods, one of which was developed with the support of IFER, are used to non-invasively and thus repeatedly examine some of the critical biochemical reactions necessary for life before, during, and after exposure to a given toxin. This model represents a significant improvement over other tissue culture models used for studying toxicity in cells, which is an alternative to using animals in testing and research.

 

1995-1996

Engelke, Dr. David R.
University of Michigan Medical School
DNA and RNA Aptamers as Replacements for Polyclonal and Monoclonal
Antibodies

 

Latham, Dr. Keith E.
Temple University, Philadelphia, PA
Construction and Development of a Mouse Embryo DNA

Objectives: “to (1) increase the number of genes that can be analyzed in a single series of samples, (2) provide other scientists with a set of dot blots with which they can determine the expression patterns of the genes in which they are interested, and (3) demonstrate the applicability of the method to the analysis of gene expression patterns at other stages of development and in embryos of other species.” The study accomplished all of its main objectives and resulted in five publications. These publications, in addition to earlier publications and publications that will follow, should lead to the adoption of this method in many more laboratories, thus reducing the number of animals used for such studies.

 

McGovern, Kathy Ann, Ph.D.
Department of Radiation Oncology, Division of Cancer Biology, University of Arizona Health Sciences Center
Development of a Versatile Tissue Model Evaluating a Cell Culture System for Toxicity Testing by NMR Spectroscopy (year 2)
During this second phase of this project, Dr. McGovern worked to complete the research project aimed at evaluating cell cultures by nuclear magnetic resonance spectroscopy.

 

1996-1997

Josephy, David, Ph.D.
Department of Chemistry and Biochemistry, University of Guelph, Guelph, Ontario, Cananda
Expression of human cytochrome P450 1A1 in Salmonella typhimurium strains, for applications in genetic toxicology testing
The objective of this project was: “the replacement of the use of animals in genetic toxicology, by further development of a new technology: expression of recombinant human enzymes in “Ames test” bacterial strains.” Genetic toxicology is the study of agents which cause heritable damage to cells. Genetic toxicology analysis is a routine component of the development of new drugs, pesticides, food additives, and other commercial chemical products. Regulatory agencies such as the FDA require that such products be demonstrated negative in these tests before their introduction into the marketplace. The researchers in this study worked to develop a new technology that obviates the need for mammalian tissue preparations in in vitro genetic toxicology studies and substitute recombinant human enzymes for rodent enzymes making the results more relevant to human health risk assessment. The researchers involved in this study noticed wider acceptance to their approach of replacing the use of animal tissues for chemical mutagenicity studies in the scientific community as their study progressed.

 

Novak, Robert, Ph.D. and Vodkin, Michael, Ph.D.
Illinois Natural History Survey, Champaign, Illinois
PCR Detection of Flaviviruses
Dr. Novak’s research for the replacement of animals for arboviral surveillance was primarily directed towards presenting an all day workshop, “PCR Detection of Flaviviruses.” The workshop was held in Norfolk, Virginia in conjunction with the annual meeting of the American Mosquito Control Association on March 24, 1996. In this workshop, Dr. Novak addressed both theoretical and applied aspects of the RT-PCR, a reverse transcription-polymerase chain reaction assay. This new technology eliminates the need to use animals and is more effective at detecting the virus that causes St. Louis encephalitis.

 

1997-1998

Josephy, David, Ph.D.
Department of Chemistry and Biochemistry, University of Guelph, Guelph, Ontario, Cananda
Replacement of Animals in Genetic Toxicology by Expression of Recombinant Human Enzymes in Ames Test Bacterial Strains (year 2)

 

Dunn, Michael G., Ph.D.
Associate Professor of Surgery and Director, Orthopaedic Research Laboratory, University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School
Human ‘tendon/ligament equivalents’ for in vitro evaluation of factors influencing tissue repair

Objectives: “(1) develop human tendon/ligament equivalents (TLEs) consisting of fibroblasts seeded onto collagen fiber scaffolds and cultured to maturity in a serum-free medium in vitro, (2) characterize normal ‘healing’ of partial and complete thickness wounds of TLEs in vitro, and (3) evaluate effects of clinically-relevant factors (e.g. pharmacological agents, mechanical loads) on TLE healing in vitro, potentially reducing or replacing currently used surgery models in animals. Tendon and ligament injuries are a major source of prolonged pain and disability in our population and thousands of animals are sacrificed each year for this research. The successful development of the TLEs could substantially reduce the use of live animals in musculoskeletal soft tissue research by providing an in vitro alternative.

 

1998-1999

Josephy, David, Ph.D.
Department of Chemistry and Biochemistry, University of Guelph, Guelph, Ontario, Cananda
Replacement of the use of animal tissues in mutagenicity assays by the engineered expression of recombinant human enzymes in bacteria (year 3)

Objective: Achieved major objective of constructing a bacterial strain which is as effective as mammalian liver preparations (“S9 fraction”) for the activation for certain chemical mutagens, making the replacement of mammalian liver preparations in genotoxicity studies possible. This project also succeeded in extending their method from the human P450 1A2 to the corresponding rat enzyme, allowing an animal-free comparison of the activities of these two species for activating mutagens. The researchers are now looking to extend the study to include a comparison of several other animals that have been used for these purposes, including mice, hamsters, and monkeys.

Van Buskirk, Dr. Robert
Department of Biological Sciences, State University of New York, Binghamton, NY.
Improving the Storage of Engineered Tissues Using Molecular Biology

 

Baust, John
State University of New York, Binghamton
Improved Cypropreservation of Human Cells and Engineered Tissues to Facilitate Product Safety Testing

Baust and his colleagues have focused on developing methods by which cells and engineered tissues can be more effectively distributed throughout the world to pharmaceutical/cosmetic companies which use in vitro techniques as a screen for product safety testing. Baust’s research lab has studied the organization of a hypothermic storage solution, HypoThermosol (HTS). IFER funding has allowed them to launch an extensive molecular biology program to further improve HTS as a storage solution.
Baust’s DNA studies focus on the molecular mechanisms underlying cell death that occur in cells stored for too long under hypothermic conditions. With IFER funding, Baust and his colleagues have found that the addition of protease inhibitors improves the performance of HTS so that it far exceeds that of VisSpan, A DuPont-Merck product used for most organ transplants and used by many in vitro toxicology laboratories for tissue slice storage. Baust’s IFER-supported work thus presents HTS as a possible, future candidate for FDA approval to support organ-transplant application as well as a preferred storage solution for the non-regulated, product safety testing market.


1999-2000

Institute for In Vitro Sciences, Inc.
Gaithersburg, MD
For support of the Institute’s efforts to develop, validate and use in vitro methodologies and to provide educational and technical resources for industry, government, trade associations and the general public.

 

Van Buskirk, Robert, Ph.D.
Department of Biological Sciences, State University of New York, Binghamton, NY.
Improving the Storage of Engineered Tissues Using Molecular Biology
Extension of grant to allow four PhD. students to continue their alternatives lab work to develop improved solutions for shipping human cells, tissues and organs for specialized research.

 

Josephy, Dr. David
Professor, Department of Chemistry and Biochemistry, University of Guelph, Guelph, Ontario, Canada
E. coli strains expressing mammalian P450 enzymes for use in genotoxicology
Objective: “Replacement of the use of animals in genetic toxicology, by expression of recombinant mammalian enzymes in E. coli bacteria; extending approach to new animal species and chemical classes.” Sucessful in constructing a bacterial strain which is as effective as mammalian liver preparations for the activation of aromative amine mutagens, prompting other researchers to develop similar systems.

 

SPREADING THE WORD

Among IFER’s earliest initiatives were the sponsorships of a series of “wet” or “hands-on” workshops and seminars which addressed state-of-the-art advances in alternatives technology.

  • 1987. IFER and the University of California, Davis co-sponsored a three day workshop that included lectures and laboratory work on isolated liver, nerve and muscle cell preparation.
  • 1988. IFER and Gibraltar Biological Laboratories (Fairfield, NJ) co-sponsored a three day workshop on alternatives in cancer research and the neutral red assay as an alternative to the Draize test.
  • 1989. IFER hosted a workshop on Computer Assisted Teaching in Denver, Co that included: interactive, 3-dimensional anatomical imaging; computer assisted surgical training; computerized case simulations for anesthetic case management; and computer modeling of the canine nervous system. The keynote speaker was Dr. Harold Modell, University of Washington and editor of Computers in Life Science Education.
  • 1990. In conjunction with the National Association of Biology Teachers, IFER
  • 1992. IFER and the American College of Veterinary Surgeons sponsored a workshop on the development and adoption of alternatives to live animals in veterinary education in Miami, Florida.

IFER has also supported, attended and/or helped to organize panels of scientists for the World Congresses on Alternatives to the Use of Animals in the Life Sciences which have been held in Baltimore (1993), Utrecht (1996), Bologna (1999), New Orleans (2002), Berlin (2005), Tokyo (2007), Rome (2009), and Montreal (2011).

 
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