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John M. Baust,
State University of New York at Binghamton (1998-2001)
Improved Cryopreservation
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 optimization 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 ViaSpan, 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 applications as well as a preferred storage
solution for the non-regulated, product safety testing
market. |
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Kathleen
Garreis, Colorado State University (1997-2000)
Research on Leishmanias
Leishmaniasis is a parasitic disease found throughout
tropical and sub-tropical areas of the world. It is transmitted
by the bite of an infected sandfly. Three manifestations
of the disease exist – the visceral form causes
enlargement of the spleen and liver and can be fatal if
not treated, the mucocutaneous form can cause severe disfigurement
to the face and the cutaneous form causes skin lesions
that can last for weeks or months. According to the World
Health Organization, approximately 12 million people are
infected with the parasite. The drugs currently used to
treat the disease can be toxic and difficult to administer,
requiring hospitalization. No vaccine has been found to
be effective in humans. New approaches to combating leishmaniasis
need to be developed and implemented.
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) which 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 Leishmania. Research is constrained
by the difficulty in performing longitudinal studies,
the variation due to the genetically heterogeneous populations
studied, the inability to experimentally manipulate the
host immune response, and the inability to perform challenge
inoculations.
Importantly, researchers are uncertain that what they
have learned about the disease in mice applies to humans
with leishmaniasis since animal models frequently do not
mimic human disease.
Garreis’s research lab found that culturing mouse
spleen cells with Leishmania 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 Leishmania.
Further, Garreis discovered the same in vitro response
to Leishmania 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
don’t 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 Leishmania. Garreis and her colleagues did not detect
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. |
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Dr.
Robert Van Buskirk, State University of New York (began
1998)
Through their IFER-supported work, Robert Van Buskirk
and his group developed instruments, engineered human
tissues, and protocols designed to facilitate the acceptance
of animal alternative techniques by pharmaceutical and
cosmetic companies so that fewer animals will be used
for product safety testing. They successfully developed
a solution called HypoThermosol that can maintain cells
and engineered tissues in a state of suspended animation
for up to a week at refrigerator temperatures so that
these products can be more widely distributed throughout
the world. After discovering that HypoThermosol can
cause cell death, they developed techniques whereby
they can stop the activation of cell death genes. |
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Dr. Michael
Dunn, University of Medicine and Dentistry of New Jersey
(began 1997)
Through his IFER-supported research, Dr. Michael Dunn
worked to develop human tendon/ligament equivalents (“TLE”)
for in vitro evaluation of factors influencing musculoskeletal
soft tissue repair. The ultimate goal of this project
was to substantially reduce the use of live animals in
musculoskeletal soft tissue research by providing an in
vitro alternative. |
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Dr. David
Engelke, University of Michigan Medical School, Department
of Biological Chemistry (began 1996)
In work supported by his IFER grant, Dr. Engelke worked
on establishing a research support facility to allow most
monoclonal and polyclonal antibody production to be replaced
by Rna aptamers. By making aptamers cheaply and at high
affinity, there would be a great reduction in the number
of laboratory animals that would need to be used. |
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Dr. Keith
Latham, Temple University, Fels Institute for Cancer Research
and Molecular Biology (began 1995)
Objectives of this project were to develop further a new
RT-PCR based method for the quantitative analysis of gene
expression in pre-implantation mouse embryos. Through
the program, Dr. Latham made significant progress in the
development of the new method and made his methodology
more widely known to other scientists. The hope is that
many of those scientists will utilize the methodology,
which would significantly reduce the number of mice embryos
necessary to conduct this sort of research. |
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Dr. Michael
Vodkin and Dr. Robert Novak (began 1994), Illinois Natural
History Survey, Department of Natural Resources
The objectives of this project were to validate recently
developed technology to reliably and sensitively detect
arboviruses in mosquito pools and to teach and disseminate
the technique to relevant diagnostic laboratories and
mosquito abatement districts thereby reducing the use
of animals as test systems. Before the development of
this technology, birds and suckling mice were primarily
being used. Validating this technology and making it more
widely known could potentially save thousands of birds
and mice each year. |
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Dr. Kathy
McGovern, University of Arizona, Cancer Biology Division
(began 1994)
Dr. McGovern developed a tissue model in which cells can
grow three dimensionally to surround an artificial capillary
bed. While studying toxicity in cultured cells has been
an obvious and frequently examined alternative to animal
studies, Dr. McGovern’s model represents a significant
improvement over tissue culture models because this model
permits examination of earlier endpoints and takes into
account cell-cell interactions. |
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