The capacity to evaluate the teratoma forming potential of differentiated embryonic stem cell populations represents an urgent need before widespread therapeutic application of these cells is possible. Researchers at Case Western Reserve University and BioInVision, Inc. will join together to create platform technologies for validating existing embryonic stem cell separation and purification paradigms (such as antibody labeling and Fluorescent Activated Cell Sorting), as well as future purification paradigms. The biologic platform consists of embryonic stem cells which report one fluorescent marker (red) when undifferentiated, and another marker (green) when differentiated down the endothelial lineage. Sorting will be evaluated in vitro by plating green cells, growing those cells in long-term culture, and determining any appearance of red undifferentiated cells. To assess cell in vivo, we will use BioInVision's unique cryo-imaging technology which can detect single fluorescent cells in a mouse. Briefly, the cryo-imaging system consists of a large stage, motorized cryostat with special features for imaging; a modified, bright-field/fluorescence microscope; and a robotic xyz imaging system positioner, all of which are fully automated by a control system to allow sectioning and tiled imaging of an entire mouse. The basic experimental paradigm will consist of sorting differentiated (green) cells and implanting them via tail injection. Days later, cryo-imaging will be used to identify teratomas and/or undifferentiated (red) cells with the potential of forming a teratoma. The cryo-imaging platform represents a distinct advantage over other imaging methods, such as bioluminescence, MRI, and PET, in that it is capable of detecting not only large overt teratomas, but also finding single cells which may have the capacity to form teratomas in the future.
Public Health Relevance: This proposal is critically concerned with the issue of public health in that its long-term goal is to provide a means for evaluating the effectiveness of embryonic stem cell selection and purification strategies prior to such cells being used therapeutically. The capacity for teratoma formation is the main barrier preventing use of embryonic stem cells clinically, for diseases such as diabetes and Parkinson's Disease and spinal cord injury.
Public Health Relevance Statement: Project Narrative This proposal is critically concerned with the issue of public health in that its long-term goal is to provide a means for evaluating the effectiveness of embryonic stem cell selection and purification strategies prior to such cells being used therapeutically. The capacity for teratoma formation is the main barrier preventing use of embryonic stem cells clinically, for diseases such as diabetes and Parkinson's Disease and spinal cord injury.
Project Terms: 2,3,7,8-Tetrachlorodibenzo-p-dioxin; ANOVA; Analysis of Variance; Anaplastic; Animal Model; Animal Models and Related Studies; Antibodies; Appearance; Automobile Driving; Bioluminescence; Blood Precursor Cell; Blood erythrocyte; Blood normocyte; Body Tissues; CAT Scan, X-Ray; CAT scan; CT X Ray; CT scan; Cell Count; Cell Differentiation; Cell Differentiation process; Cell Isolation; Cell Number; Cell Segregation; Cell Separation; Cell Separation Technology; Cells; Color; Computed Tomography; Computer Programs; Computer software; Computerized Axial Tomography (Computerized Tomography); Computerized Tomography, X-Ray; Contracting Opportunities; Contracts; Data; Detection; Diabetes Mellitus; Dibenzo(b,e)(1,4)dioxin, 2,3,7,8-tetrachloro-; Disease; Disorder; Dose; Drivings, Automobile; DsRed; Dysembryoma; EMI scan; ES cell; Effectiveness; Embryo; Embryonic; Endothelial Cells; Erythrocytes; Erythrocytic; Event; F element; FP593; Fee-for-Service Plans; Fees for Service; Fluorescence; Fluorine; Frequencies (time pattern); Frequency; Future; GFP; Glossary; Goals; Green Fluorescent Proteins; Hematopoietic stem cells; Histology; Idiopathic Parkinson Disease; Image; Image Analyses; Image Analysis; Imagery; Imaging technology; Implant; In Vitro; Injection of therapeutic agent; Injections; Instrumentation, Other; International; Intestinal; Intestines; Investigators; Isotopes; Joint Ventures; Label; Laboratories; Lewy Body Parkinson Disease; Licensing; Location; MR Imaging; MR Tomography; MRI; Magnetic Resonance Imaging; Magnetic Resonance Imaging Scan; Magnetism; Mammals, Mice; Manuals; Marrow erythrocyte; Medical Imaging, Magnetic Resonance / Nuclear Magnetic Resonance; Medical Imaging, Positron Emission Tomography; Medicine; Methods; Methods and Techniques; Methods, Other; Mice; Molecular; Murine; Mus; Muscle, Involuntary; Muscle, Smooth; Myelopathy, Traumatic; NMR Imaging; NMR Tomography; Nuclear Magnetic Resonance Imaging; Organ; PET; PET Scan; PET imaging; PETSCAN; PETT; Palpable; Paralysis Agitans; Parkinson; Parkinson Disease; Parkinson's; Parkinson's disease; Parkinsons disease; Phase; Phenotype; Photons; Population; Position; Positioning Attribute; Positron Emission Tomography Scan; Positron-Emission Tomography; Prevention; Primary Parkinsonism; Probability; Progenitor Cells, Hematopoietic; Promoter; Promoters (Genetics); Promotor; Promotor (Genetics); Proteins; Proton Magnetic Resonance Spectroscopic Imaging; Public Health; ROC Analysis; Rad.-PET; Red Blood Cells; Red Cell; Red blood corpuscule; Red cell of marrow; Reporter; Reporter Genes; Reporting; Research; Research Contracts; Research Personnel; Researchers; Reticuloendothelial System, Erythrocytes; Risk; Risk Estimate; Robotics; Scheme; Science of Medicine; Screening procedure; Smooth muscle (tissue); Societies; Software; Sorting - Cell Movement; Spinal Cord Trauma; Spinal Trauma; Spinal cord injured; Spinal cord injuries; Spinal cord injury; Staging; System; System, LOINC Axis 4; TCDD; Tail; Techniques; Technology; Technology Assessment; Teratoid Tumor; Teratoma; Testing; Tetrachlorodibenzodioxin; Therapeutic; Time; Tissues; Tomodensitometry; Tomography, Xray Computed; UV laboratory microscope; Ultraviolet Microscopes; Undifferentiated; Universities; Variance Analyses; Veins; Visualization; X-Ray Computed Tomography; Zeugmatography; base; blood corpuscles; bowel; catscan; cell imaging; cell sorting; cellular imaging; commercialization; computed axial tomography; computer program/software; computerized axial tomography; computerized tomography; cryostat; diabetes; disease/disorder; drFP583; driving; ds red protein; dsFP593; embryonic stem cell; fluorescence microscope; fluorescence/UV microscope; fluorescent microscope; gamma Actin; gene product; genetically modified cells; image evaluation; imaging; imaging modality; improved; in vivo; instrumentation; laboratory fluorescence light microscope; magnetic; model organism; molecular imaging; palpable disease; prevent; preventing; progenitor; public health medicine (field); public health relevance; red fluorescent protein; screening; screenings; sorting; stem cell of embryonic origin; stem cell population; stem cell therapy; tomography; tool; urinary
