Animal Sciences presentations (MU)
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Items in this collection are public presentations made by Division of Animal Sciences faculty, staff, and students, either alone or as co-authors, and which may or may not have been published in an alternate format. Items may contain more than one file type.
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Item Commercialization of novel biomarkers of male fertility in humans and farm animals(2010-03) Sutovsky, Peter; University of Missouri (System); Missouri Life Sciences Summit (2010: University of Missouri--Kansas City)Each year, US infertility clinics treat 135,000 couples who fail to conceive naturally. Up to 40% of these infertility cases can be attributed to male infertility stemming from poor sperm quality. An additional 20% of couples present at the clinic with idiopathic, unexplained infertility, some of which is in fact hidden, misdiagnosed male infertility. Due to a paucity of accurate diagnostic methods and efficient treatments, the success rate of assisted fertilization, measured by live births, stagnates around the disappointing 35% margin. Many parallels exist between human male infertility and male reproductive performance in farm animals, where the estimated losses from inferior reproductive performance amount to millions of dollars annually, according to USDA.Item Swine models of Human Conditions(2010-03) Wells, Kevin D.; University of Missouri (System); Missouri Life Sciences Summit (2010: University of Missouri--Kansas City)Since the introduction of Dolly, the first cloned sheep, genetic engineering of livestock has advanced to the point that nearly any genetic modification appears feasible. Genes can be introduced, deleted or altered in cultured cells. These cells can be evaluated in culture and then used to regenerate live animals by somatic cell nuclear transfer (cloning). Since pigs share similar physiology with humans and have proven to be excellent models for many human conditions, genetic engineering and animal cloning are being used to produce models of human disease states and disorders. These models are proving to be particularly important for situations where laboratory rodents have proven to be inadequate. Current successes of genetically engineered swine biomedical models will be presented.Item Porcine induced pluripotent stem cells (piPSC) for expanding the use of swine in biomedical research(2010-03) Ezashi, Toshihiko; Telugu, Bhanu Prakash V. L., 1977-; Roberts, R. M. (Robert Michael), 1940-; University of Missouri (System); Missouri Life Sciences Summit (2010: University of Missouri--Kansas City)Our goal is to create porcine pluripotent stem cells, i.e. ones capable of differentiating into all cell types of the body that can expand the use of swine as a biomedical model for studying human disease. It is well established that mouse embryonic stem cells (ESC) are an excellent source of material for successful cloning and for incorporation into chimeras. However, the establishment of porcine ESC from the embryos has proven to be elusive. There has been a similar lack of success with other ungulate species. Establishing a technology for deriving induced pluripotent stem cells (iPSC) from farm animals will allow the gene knock-in/knock-out methods that have revolutionized mouse genetics to be applied to farm species. Importantly pig is a potentially useful model for studying human pathologies due to similarities in organ size, immunology and whole animal physiology between the two species. If the safety and efficacy of stem cell transplantation is to be tested in an animal model before being applied to humans, the pig would likely be a species of choice. The ability to derive porcine (p) iPSC lines from a particular outbred animal and conduct tissue transplantation on the same pig later and follow the success of the transplant over the course of months or even years would be a particularly valuable advance. Additionally the ability to provide piPSC from animals with valuable traits would provide a permanent source of cells for clonal propagation that would likely avoid the inefficiencies and problems arising from somatic cell nuclear transfer (SCNT), where the vast majority of cloned offspring die or are developmentally abnormal. We have created piPSC from embryonic fibroblasts and umbilical cord mesenchyme by a similar strategy used for the mouse and human, namely ectopically expressing reprogramming genes in somatic cells. The piPSC resemble human ESC, express the typical gene and surface antigen markers of ESC, proliferate continuously in culture, possess high telomerase activity, form embryoid bodies, and differentiate along the three main germ line lineages. Our aim is to demonstrate that piPSC can be directed to differentiate along defined lineages, specifically towards neuronal tissue, hematopoietic lineages and various mesoderm derivatives including cardiomyocytes by using protocols based on those used successfully with human and murine ESC. These experiments will allow such cells to be used for tissue grafts that are matched genetically to recipients and tested for their safety in transplantation. We shall also establish parameters for routine gene targeting in piPSC, with the ultimate goal of creating genetic models for human diseases where mouse models are inappropriate. In summary, the piPSC lines developed will have enormous utility for exploiting the pig as a model in human pre-clinical applications. Supported by Missouri Life Sciences Board Grant 00022147 and NIH grant HD21896Item Derivation and characterization of LIF and FGF2 dependent Porcine Induced Pluripotent Stem Cells [abstract](2010-02) Telugu, Bhanu Prakash V. L., 1977-; Ezashi, Toshihiko; Roberts, R. M. (Robert Michael), 1940-; University of Missouri (System); Missouri Life Sciences Summit (2010: University of Missouri--Kansas City)Authentic embryonic stem cells (ESC) have probably never been derived from the inner cell mass (ICM) of pig, despite over 25 years of effort. Recently, several groups, including ours, have reported derivation of induced pluripotent stem cells (iPSC) from swine by reprogramming somatic cells with a combination of four factors (OCT4, SOX2, KLF4, C-MYC) delivered by retroviral transduction. The piPSC resembled FGF2- dependent human (h) ESC and are likely to advance swine as a model in biomedical research, since grafts could potentially be matched to the animal that donated the cells for re-programming. The dependence of piPSC on FGF2 also draws parallels to murine pluripotent stem cells derived from advanced epiblast, so-called 'epiblast stem cells'. Indeed, an emerging concept is that there are two kinds of ESC, one dependent on FGF2 and not competent to contribute to germ-line chimeras, the other upon LIF and germ-line competent. The objective of our recent investigations has been to develop LIF- dependent piPSC by using the same reprogramming factors but selecting the colonies on a modified LIF- medium supplemented with two kinase inhibitors, CHIR99021, which inhibits GSK-3beta, and kenpaullone, which inhibits both GSK-3beta and CDK1. The LIF-dependent piPSC, derived here from outgrowths of umbilical cord mesenchyme, expressed markers consistent with pluripotency and bore a striking resemblance to ICM-derived murine ESC in colony morphology, culture characteristics, and short cell cycle time. Currently, the ability of LIF-piPSC to give rise to teratoma and chimeras is under investigation. Supported by Missouri Life Sciences Board Grant 00022147 and NIH grant HD21896.Item Swab Test Using ELISA Technique for Diagnosis of Brown Recluse Bites(2010-03) Stoecker, William V.; Black, Elizabeth; Stricklin, Sherea; Green, Jonathan A., 1969-; University of Missouri (System); Missouri Life Sciences Summit (2010: University of Missouri--Kansas City)Envenomations by the brown recluse spider, Loxosceles reclusa, are a significant source of morbidity and occasional mortality in endemic regions of the United States, and misdiagnoses are common. A survey of physicians in the endemic area has shown the economic viability of an accurate diagnostic test for these spider bites. Development and testing of an optimized loxosceles venom test kit will present significant challenges. Unlike the routine construction of ELISAs dedicated to the detection of a single protein, the proposed kit immunoassay will detect venom containing multiple proteins, including a unique physiologically active protein-sphingomyelinase D (SMD) abundantly present in the venom. In preliminary and Phase I research, our polyclonal assay has shown good in-vitro and in-vivo sensitivity and specificity. Our research shows that identifiable amounts of venom in clinical envenomations persist for at least 5 days, our longest probable envenomation, which covers the majority of cases encountered in emergency rooms and clinics. Phase I testing showed high specificity, with none of 60 competing inflammatory diagnoses showing reactivity above background. The limits of sensitivity, in-vivo specificity, and the duration of detection of the recently developed venom-affinity purified test are unknown. Phase I development of a venom-affinity purified polyclonal assay has successfully allowed several orders of magnitude increase in sensitivity above that of other assays. Remarkably, the current swab test now operates in the femtogram range (one billionth of a microgram). This allows detection of venom even after the victim has washed the suspected bite site. Phase I analysis determined that monoclonal antibodies raised in a bioreactor had relatively weak venom affinity and provided no improvement over the polyclonal antibody-based assay. If conjugated monoclonal techniques fail in Phase II, then the assay will use only polyclonal antibodies in an optimized microtiter plate assay. A prototype microtiter colorimetric assay is now available and ready for testing by hospitals with ELISA reader capability. A simplified tube kit test or other format developed during Phase II will allow testing by small hospitals. These clinical studies will allow determination of sensitivity and specificity of our test, with the goal of FDA device approval and ultimately marketing. Clinical application of an optimized assay would save the morbidity and expense due to inappropriate diagnosis and treatment of various skin conditions with presentations similar to Loxosceles envenomations. Techniques used in the successful detection of this spider venom are directly applicable to bites from S. American Loxosceles species, which are responsible for additional deaths each year. The swab venom assay technique could be applicable to envenomations by spiders outside the Loxosceles genus.