Core A (Imaging) is vital for the MRI imaging of fetuses in utero and teratomas, and ultimately for trackingtransplanted stem cells. In vitro real-time and laser-scanning confocal, quantitative FRAP, wide-field and electronmicroscopy are also provided to all three subprojects to evaluate the dynamics of live cell indicators of pluripotency inhESC, nhpESC and PG cells in order to better define characteristics of the pluripotent state. The imaging core providesexceptional high-sensitivity detection of stem cell dynamics in vitro, in utero and in vivo, essential for each of theresearch projects. To ensure maximum utility of the Imaging Core, we will combine the strengths of in vivo noninvasiveMRI with in vitro imaging by conventional, confocal, and electron microscopy. At the macroscopic wholeanimal level, we will utilize new small animal imaging technologies, such as MRI. The primary roles of the ImagingCore will be to: perform longitudinal MRI studies of pregnant monkeys and monitor viability and phenotypicdifferences of the chimeric fetus and placenta throughout pregnancy; employ high-resolution 3D MRI of fixed and liveES-derived NHP embryos to examine stem cell contributions; and monitor teratoma formation in vivo.
Aim 1. The imaging core will provide total body non-invasive functional imaging to all three projects. The core isresponsible for tracking cell lineage contributions in ES and EG chimeric embryos and the fates of transplanted hEScells. The core will also use high resolution MRI to map fetal development and teratoma formation, and performmagnetic resonance microscopy on excised teratomas prior to sectioning. This core will also work towards developingand optimizing techniques for the tracking of stem cells in vivo including SPIO labeling of ES and EG cells for MRI.
Aim 2. The imaging core will provide instrumentation, oversight and guidance for fixed, immunocytochemistry, livecell confocal, real-time dynamic and electron microscopy for assaying cell structure and dynamics in pluripotent anddifferentiated human embryonic stem cells, especially live and fixed pluripotent marker assays to better characterizeundifferentiated human and nonhuman primate ES and EG cells. This core will also perform fundamental studies of ESand EG cell contributions to chimeric embryos and imprinting status of ES and EG cells, as well asimmunocytochemistry and immunohistochemistry of teratomas and embryoid bodies.
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