Lee, Jaewon Human gut inflammation-on-a-chip for replacement of animal models University of Texas at Austin Mentor: Dr. Hyun Jung Kim
2015 - 2017
Graduate student Jaewon Lee, in the Department of Biomedical Engineering at the University of Texas at Austin, has received a fellowship for her project which aims to study intestinal inflammation using the human gut-on-a-chip microphysiological system. Dozens of animal models of gut inflammation exist but have limitations including expense, time, ineffectiveness, and ethical challenges that can be overcome with this sophisticated, human-relevant model. In this proposal, Jaewon plans to co-culture intestinal cells, gut microbes and immune cells in a sophisticated human intestinal microphysiological system and examine the effect of probiotic and prebiotic therapies after inflammation is induced, in an effort to reduce and replace animal use in this area.
Tan, Sun Nee Structural and Functional Neuroplasticity of Parkinson’s Disease Following a Sensori-motor Contingent Musical Walking Intervention (Ambulosono) University of British Columbia Mentor: Dr. Martin McKeown
2014 - 2016
Sun Nee Tan, a Ph.D. candidate in the Graduate Program in Neuroscience at the University of British Columbia, is a new recipient of the IFER fellowship. Under the mentorship of Dr. Martin McKeown, Sun Nee will be using state-of-the-art brain imaging techniques on human subjects for her project, “Structural and functional neuroplasticity of Parkinson’s disease following a sensori-motor contingent musical walking intervention (Ambulosono).” Her study will determine the effectiveness of a music-contingent walking exercise program as an intervention for Parkinson’s disease subjects. While many animal models are relied upon heavily in neuroscience studies, despite their limited translatability to human populations, using imaging techniques on human subjects to study neuroplasticity may replace conventional research which uses animals for this purpose.
Hassell, Bryan Human Cancer-on-a-Chip as a Replacement for Animal Testing Wyss Institute for Biologically Inspired Engineering at Harvard University Mentor: Dr. Donald Ingber
2014 - 2016
Bryan Hassell, a Ph.D. candidate working in Dr. Donald Ingber’s laboratory at the Wyss Institute for Biologically Inspired Engineering at Harvard University, is a new Graduate Fellowship recipient. Bryan’s project, “Human cancer-on-a-chip as a replacement for animal testing” seeks to develop an organ-on-a-chip platform to determine how lung cancer cells respond to chemotherapy depending on their organ-specific microenvironment and to identify new anticancer therapies. Bryan’s physics and engineering background, in combination with the extensive biological training he is receiving in the Ingber lab, will allow him to take a multidisciplinary approach toward developing an innovative model that has the potential to replace the use of animals.
Schlesinger, Erica In Vitro 3D Flow Through System for Improved Intestinal Permeability Model
University of California, San Francisco Mentor: Dr. Tejal Desai
2014 - 2015
Erica Schlesinger, a Ph.D. student at the University of California San Francisco working in Dr. Tejal Desai’s lab, is a new IFER grant recipient. Her project, “In vitro 3D flow through system for improved intestinal permeability model” aims to develop a model in which human intestinal cells are grown on a tubular scaffold to better mimic the microenvironment of the intestine. Fluid, containing experimental drugs, can be flowed through the system, and drug absorption can be assessed. This new model has the potential to improve upon existing in vitro approaches to study drug absorption and may reduce the use of animals as well.
Tran, David Development of a Patient-Specific High-Throughput Cardiac Drug-Screening Platform
University of California, Irvine Mentor: Dr. Steven George
2013 - 2015
David Tran, a Ph.D. student working under Dr. Steven George at the University of California, Irvine, has received a second year of IFER support for his project, “Development of a patient-specific high-throughput cardiac drug-screening platform.” David has been generating an in vitro model using human induced pluripotent stem cells as a source for human cardiomyocytes to examine the pharmacokinetics and toxicity of drugs. Over the past year, his model has undergone several changes to improve its functionality and enable it to better characterize the effects of drugs on heart function. Changes include the use of cells that express an ultrasensitive calcium indicator protein and the incorporation of electrodes on the bottom surface of the cellular chamber to better monitor changes in cardiac tissue electrophysiology. Development of such a model is critical, as a significant portion of rejected drugs fail in humans because of cardiotoxic side effects not evident in animal models.
Sparks, Nicole In Vitro Prediction of Skeletal Teratogenicity of Environmental Chemicals
University of California, Riverside Mentor: Dr. Nicole l. zur Nieden
2012 - 2015
Nicole Sparks, a Ph.D. student under the mentorship of Dr. Nicole l. zur Nieden at the University of California, Riverside, is in her third year of funding from IFER for her project “In vitro prediction of skeletal teratogenicity of environmental chemicals.” Nicole continues to make impressive progress in her efforts to develop a method to accurately predict and identify compounds that can cause skeletal abnormalities during development using human stem cells. As existing teratogenicity tests rely on animals and have low predictively for human effects, the need for a human-relevant in vitro embryotoxicity screen has never been greater.
Meyerett, Crystal Development of a Cell Culture – based In Vitro Prion Amplification Assay as a Diagnostic Tool for Chronic Wasting Disease
Colorado State University Mentor: Mark Zabel
Diagnosing prion diseases such as the so-called “mad cow” disease in cattle, scrapie in sheep or chronic wasting disease (CWD) in deer, elk and moose, relies on either transmitting the disease agent, know as prions, to live animals or detection of a surrogate marker for disease. Recently, a major breakthrough in prion diagnostics known as Protein Misfolding Cyclic Amplification (PMCA), conceptually similar to DNA PCR, has enabled preclinical, antemortem prion detection, but requires the use of animal products. We propose to develop a cell culture-based PMCA assay to detect prions that abrogates the need for live animals or animal products.
Ho, Hsiang Development of an Entirely Human Derived Skin Equivalent Model that will Replace Animal Models for In Vitro Testing of Melanoma Chemopreventive Agents
University of California - Irvine Mentor: Anand Ganesan
2009 - 2010
Animal models for validating melanoma chemopreventive agents are unavailable. In this proposal, we describe the development of a human skin equivalent model for chemopreventive agent validation. Loss of function analysis will be utilized to validate that known genes that inhibit (E-cadherin) or promote (N-cadherin) melanoma formation impact tumor progression in our tissue model. Real time multiphoton microscopy will be utilized to measure the impact of individual genes or chemopreventive agents on the behavior of normal or neoplastic melanocytes In Vivo. Eventually, this skin equivalent model will obviate the need for laboratory animals in preclinical testing of melanoma chemopreventive agents.
Remais, Justin Development of an Animal Replacement Assay for the Detection of
University of California, Berkeley, CA Mentor: Robert C. Spear
2007, 2009 - 2010
The parasitic disease schistosomiasis infects 200 million people in the developing world, including more than 800,000 in China. The parasite’s infective stage, called a cercaria, inhabits irrigation canals and drainage ditches, which is where people become infected. Detecting and isolating cercarial contaminated waters has been shown to be vital for protecting human health. The current practice in China uses sentinel mice, a slow and expensive method that exhibits poor spatial and temporal resolution and costs the lives of approximately 10,000 mice per year in China alone. In response to these limitations, we aim to develop a molecular detection technique designed to improve the resolution of cercarial risk measurements at less expense and without the use of sentinel animals.
Dean, Dylan Microtissues in Micromolded Hydrogels: Reducing Animal Use With In Vitro Tissue Analogues
Brown University Mentor: Jeffrey Morgan
2007 - 2008
Pharmaceutics is a multi-billion dollar industry investing heavily in two-dimensional cell culture and expensive, inefficient and often unnecessary animal research. We have developed a novel, high throughput technology for generating micro-tissues of prescribed geometry in non-adhesive, agarose hydrogels micro-molded from sterile casts of wax models designed in CAD. These hydrogels will be used first to investigate intracellular contributions to self-assembly, then to investigate the applicability of the technology as a replacement for animal research in clinical or pharmacological trails. Further, the technology will be actively shared with both scientific and commercial communities for the promotion of more responsible research.
Arora, Deepika In Vitro Cell Monolayer-based Assay for Lung Epithelial Inflammation and Barrier Damage: Their Induction by Cigarette Smoke and Recovery By Pharmaceutical Aerosols
Virginia Commonwealth University Mentor: Masahiro Sakagami
The goal of this graduate project is to develop a unique in vitro “cell monolayer” –based assay system to study lung epithelial inflammation and barrier damage in response to cigarette smokes and pharmaceutical aerosols. Air-interface cultured and confluent monolayers of human lung epithelial cells, Calu-3, will be used, alongside transient transfection with an inflammation-related transcription factor, NFкB. Specifically, the project will pursue the following two aims: 1. To characterize induced epithelial inflammation and barrier nature of the Calu-3 monolayers in response to cigarette smoke deposition; 2. To assess anti-inflammatory actions of inhaled corticosteroids upon their aerosol deposition on the Calu-3 monolayers.
Mallampati, Ramya Evaluation of Epiderm FT-200 (EFT-200) culture as an In Vitro model for skin irritation
Florida Agricultural and Mechanical University Mentor: Mandip Sachdeva
2006, 2008 - 2009
Skin irritation can be defined as a non-specific inflammatory response to chemical damage which is characterized by erythema, mild edema and scaling (Irritant contact dermatitis). Understanding the signaling mechanisms involved in chemical induced irritation is recognized to be of great significance and several animal studies have been reported. However, very few studies have been performed with human skin cultures. The objective of our research is to use Epiderm full thickness ski (EPI-FT200) as alternative in vitro model for skin irritation studies. Jet fuel (JP-8) and its major components (Nonane, dodecane, tridecane, tetradecane, hexadecane) will be used as model irritant chemicals.
Kim, Sean H.J. Development of a Multi-Scale, Multi-Aspect Framework and Models for In Silico Experimentation of Mammalian Organ Systems
University of California, San Francisco and Berkeley Mentor: C. Anthony Hunt
2006, 2007, 2008
Our project aims to develop and thereby demonstrate the feasibility of a new class of multi-scale, multi-aspect in silico models and an experimentation framework. The models will be prototypes designed to simulate aspects of mammalian organ systems. The research is early stage. The new class of models is expected to progressively evolve into viable alternatives to the animal derived in vitro and in vivo models used in research (especially pharmaceutical) and education. This project includes development and validation of two early stage models: the blood-brain barrier and the functional unit of the kidney.
Calabro, Anthony R. Effects of Trace Metals on Tight Junction Proteins in Cultured Caco-2 and Mouse Embryonic Stem Cells
St. John’s University, New York Mentor: Frank A. Barile
2005 - 2006, 2007 – 2008
One function of epithelial cells in multicellular organisms is their ability to regulate the transport of molecules across the apical and basolateral surfaces of cellular environments. Proteins that form tight junctions (TJs) create connections between adjacent cells that alter the passage of ions and other molecules. Three families of proteins constitute TJs; claudins, occludins and junctional adhesion molecules (JAM), all of which are associated with actins – that is, the cell anchoring proteins. Consequently, this in vitro study has two objectives: 1. to develop a cell culture model, using immortal human colon cells and mouse embryonic stem (mES) cells, to determine acute cytotoxicity of trace metals in vitro, and; 2. to characterize the effects of the trace metals on the expression of TJ proteins in the cell culture models.
Haubner, Aaron Design and Discovery of Novel Subtype-selective Nicotinic Receptor Agonists and Antagonists Utilizing Animal-free Cell Culture and Microplate Technologies
University of Kentucky, Lexington, KY Mentor: Peter A. Crooks
2003 – 2004, 2005
This project will utilize cloned human cell lines that express a variety o nicotine receptors to pharmacologically evaluate and determine the mechanism of potential drg candidates. The goal is to establish experimental protocols that will eventually replace the use of animals in nicotine drug activities and mechanism screens, as well as improve experimental assay precision and predictability.
Mayo, Susan L. Oral Immunization of Chickens for Refinement of Egg-Yolk Antibody Production: Implementation of Four R’s in Antibody Production
Uppsala University, Sweden Mentor: Jann Hau
2003 - 2004, 2005
We have demonstrated that chickens are excellent antibody producers utilizing antibodies purified from egg-yolk instead of blood from mammals. This project will develop lenient oral immunization techniques so polyclonal antibody production may cease to entail any unpleasantness for the animal. We achieve: Refinement because animals are not restrained, injected or blood sampled; Reduction because chickens produce far more antibody in the egg-yolk than can be obtained from mammals of comparable sizes; Replacement because antibody production will no longer be an unpleasant procedure. Finally, Responsibility is demonstrated by transforming antibody production into a normal animal friendly farm animal production system.
Zhao, Bin Development of a Human Kidney Cell Line Model for the Assessment of Antiviral Drug Transport
St. John’s University, New York Mentor: Sue M. Ford
This project will determine if and how antiviral drugs can be transported in cultured human cells. This project will eliminate the use of animal cells for transport studies and potentially replace monkeys in pre-clinical studies of kidney toxicity.
This project will give Ms. Bin Zhao, MS candidate, experience in characterizing a new human kidney cell culture model for pre-clinical assessment of therapeutic agents. As a pharmacology student she will learn cell culture techniques applicable to her discipline, rather than whole animal or ex vivo methods. The human kidney cell line will be grown on filters and examined for transport characteristics. Initially validation will be done with prototype compounds followed by testing with antivirals. The model she helps develop will be valuable to pharmaceutical companies in reducing and replacing animals in the pre-clinical testing of antiviral drugs.
Suuronen, Erik Development of an Innervated Functional Human Corneal Equivalent for In Vitro Testing
University of Ottawa, Canada Mentor: May Griffith
2001, 2002 – 2003
We recently developed a human corneal equivalent using tissue engineering methods and human corneal cell lines as a possible replacement for animal ocular toxicity testing. Although the corneal equivalent gives quantifiable changes in transparency in response to different chemicals that indicate different degrees of cellular damage, it cannot provide indications of sensitivity and pain. Our goal, therefore, is to develop a more complete, fully innervated corneal model that can also predict of responses for stimuli such as pain. This could, in the future, provide a superior alternative to the use of animals in testing ocular irritancy and chemical toxicity.
Thompson, Hillary, Masters level Graduate Student Project
The Zoo as a Natural Learning Laboratory: Developing a Behavior Research and Training Unit with ISU and Miller Park Zoo
Illinois State University Mentor: Valeri Farmer-Dougan and Thomas Critchfield
2001 – 2002, 2003
This project uses a zoo as an alternative to traditional animal laboratories. Zoo animals need enrichment and training, and psychology students need opportunities to develop training and observation skills traditionally obtained in animal laboratories. The zoo thus provides an ethical, cost-efficient alternative. It reduces the number of laboratory animals used for training, and provides stimulation to the zoo animals. Public attitudes may also be impacted: Public education components are included on animal behavior, appropriate use of research animals, and animal welfare. A program evaluation assesses the impact on student training, along with changes in knowledge of and attitudes toward animal welfare.
Dloniak, Stephanie Marie Evaluation of Hormonal Status, Parasite Load, and Behavioral Correlates in Free-ranging Spotted Hyenas (Crocuta crocuta)
Michigan State University Mentor: Kay E. Holekamp, Ph.D. in collaboration with Dr. Steve Glickman, University of California - Berkeley
2000, 2001, 2002
This proposal describes a three-year graduate project concerned with behavioral endocrinology of free-ranging spotted hyenas, Crocutacrocuta. The project has three main goals, which are to 1) validate various steroid hormone assays for use with spotted hyenas, 2) use these assays to evaluate hormonal status in free-ranging spotted hyenas using over 7 years of archived fecal samples, and 3) examine the relationships between hormonal status, fecal parasite loads, and agonistic behavior. The proposed non-invasive research will yield rare and valuable comparative data revealing how physiological factors interact to shape mammalian behavior, and will enhance understanding of carnivore behavior and endocrinology to facilitate decision making by wildlife conservation managers.
Schuppli, Catherine Governance of Experimental Animal Use in Canada: The Effectiveness in Reaching Ethical Goals
University of British Columbia Mentor: David Fraser
2001, 2002, 2003
In Canada, the Canadian Council on Animal Care was created to uphold ethical standards of replacement, reduction and refinement of animal use in research, product testing and education. However, the effectiveness of this system in achieving these goals has never been formally evaluated. Decisions about experimental animal use are influenced by several levels of governance (the local Animal Care and Use Committee, the national inspection program, policy documents), as well as external factors such as regulator requirements, granting bodies and industry. This study will examine these relationships to evaluate the effectiveness of the system in achieving ethical goals. This is a case study that can be used as a template for similar evaluation in other countries.
Shunick, Margaret Feelings, Beliefs and Values About Animals in the Wild
University of Montana Mentor: Deni Elliott
1999, 2000, 2001 – 2002
Medical researchers address animal welfare concerns using guidelines known as the Three R’s (expanded by IFER to the Four R’s). Wildlife biologists, animal scientist, and animal protection advocates have no similar methodology
for assessing the well-being of wild species. This project anticipates finding subjective agreement regarding animal welfare in the debate over Yellowstone National Park’s bison; concerns for animal well-being are expected to provide common ground for people divided over management issues. This project’s subsequent analysis of land use and animal regulation will use animal welfare defined by the Four R’s as assessment criteria for the recommendation of policy changes.
The application of a human/animal empathy scale developed by Project Director Marianne Spitzform, the “Interpersonal Reactivity Index Animal Form,” (also funded in part by IFER) will be used to gauge the degree of operational empathy that various segments of the test group exhibit towards wildlife.
Cocco, Regina and Lukovic, Dunja Distinct Modes of Macrophage Recognition for Apoptotic and Necrotic Cells Are Not Specified Exclusively by Phosphatidyleserine Exposure
University of Illinois Mentor: David Ucker
1999 - 2000, 2001 – 2002
While examining cell death is a vital part of the morphological shaping of tissues in development, this project focuses on the properties unique to the dying cells that determine the mode and outcome of phagocytic clearance of the dead cells from tissue.
Cell Death is critical in normal organismal development and homeostasis, particularly for shaping and maintaining appropriate cellular networks. We have addressed previously the fundamental question of whether a common cell-autonomous effector mechanism pertains in distinct cases of cell death. That work has led to the identification of a thematically conserved, ordered pathway for cellular destruction.
The ability of a dying cell to trigger phagocytosis without eliciting an inflammatory response likely is the overriding biological purpose of the physiological cell death process. The fundamental question in this context is how recognition without inflammatory response is assured. We have developed a novel In Vitro system with which to study this issue. We will dissect events that lead to appropriate recognition and clearance by phagocytic cells. Our quantitative methodology, which relies on clonal populations of established cell lines, offers significant scientific and technical advantages for this work, as well as eliminating the need for whole animal studies or the use of animals as a source of experimental cell populations.
As understanding of the regulation, mechanism, and outcome of the physiological cell death process will offer insights to normal cell and tissue development, and may provide new views of aging and treatments for pathological conditions, including autoimmune diseases, neurological degeneration, and cancers.
Baust, John M. Cell Viability Improves Following Inhibition of Cryopreservation-Induced Apoptosis
State University of New York - Binghamton, N.Y. Mentor: Robert Van Buskirk
1998, 1999, 2000
Traditional approaches to cryopreservation of human cells and engineered tissues rely on the addition of molar concentrations of penetrating cryoprotectants contained in either isotonic or hypotonic solutions. This approach rarely considers the ionic balance, buffering capacity or other factors thought to be necessary to avoid hypothermic stress. John Baust’s research in the past few months has shown that HypoThermosol is a superior cryoprotectant to the industry standard. More specifically, cells separately cryopreserved
(-196 deg. C ) in either HypoThermosol or 5% DMSO in cell culture media yielded a 30% survival rate; wheras cells cryopreserved in HypoThermosol +5% DMSO had up to a 82% survival rate. These observations suggest that HypoThermosol may be both a superior hypothermic solution
(storage at 4 deg C ) as well as a superior cryoprotectant
(storage at -196 deg C).
Under IFER funding John Baust plans to conduct the following studies: 1. Test the addition of selected HypoThermosol additives listed in ”Laboratory Research Background” to determine if these additives improve HypoThermosol’s ability to act as cryoprotectant. 2. Test optimized HypoThermosol as a cryoprotectant for the most common cells strains used for product safety testing ( normal human epidermal keratinocytes, human renal cells, human hepatocytes). 3. Isolate the role of apoptosis and necrosis in cryopreservation. 4. Test the optimized HypoThermosol solution for its ability to cryopreserve the engineered human epidermis, EpiDerm (MatTek Corporation, Ashland, MA). The sum consequence of John Baust’s research will be to develop improved techniques for cryopreservation to facilitate the distribution of engineered human cells and tissues for product safety testing.
Garreis, Kathleen A. Leishmaniasis Disease and It’s Effect on Humans
Colorado State University Mentor: Richard Titus
Leishmaniasis is a parasitic disease found throughout tropical and sub-tropical areas of the world that affects approximately 12 million people according to World Health Organization estimates. Extensive work in the mouse model for leishmaniasis showed that resistance to the parasite is mediated by white blood cells known as Th1 T cells. There are other white blood cells (Th2 cells) that cause disease progression. This same Th1/Th2 phenomenon is now known to occur in tuberculosis and AIDS. Therefore, leishmaniasis has become a widely studied model since it provides insights into many of the world’s most serious diseases. Unfortunately, little is known about the disease process in humans infected with leishmaniasis since animal models frequently do not mimic human disease.
Garreis’s research lab found that cultivating mouse spleen cells with Leishmaniasis parasites resulted in the activation and proliferation of the Th1 and Th2 cells that are activated in mice with the disease. This discovery reduced the number of mice used in their lab and due to the controlled nature of in vitro assay systems, they obtained information that would be impossible to obtain using mice infected with leishmaniasis. Garreis also discovered the same in vitro response to Leishmaniasis occurs using peripheral blood mononuclear cells (PBMC) from human donors. This method eliminates the limitations of past research on humans and on animal models. The studies can be fully controlled and since the same donors can be repeatedly tested, their studies do not suffer from genetic variation. Garreis found that some donors responded with a strong T1-like (protective) response while others had a weak response. Results also showed differences between the human and mouse response to Leishmaniasis. Garreis and her colleagues did not detecte Th2 cell development in PBMC from human donors as they did in mice.
Using PBMC from human donors, Garreis has replaced the mouse as the major research tool for Leishmaniasis and sped up the development of vaccines for human Leishmaniasis and other diseases such as AIDS and tuberculosis.