In Vivo Assessment of Embryonic Stem Cell Teratoma Prevention

胚胎干细胞畸胎瘤预防的体内评估

• 批准号: 7804019
• 负责人: Horst A. von Recum
• 金额: $ 24.83万
• 依托单位: BIOINVISION, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-15 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7804019
• 关键词: Analysis of Variance; Animal Model; Antibodies; Appearance; Automobile Driving; Bioluminescence; Cell Count; Cell Differentiation process; Cell Separation; Cells; Color; Computer software; Contracts; Data; Detection; Diabetes Mellitus; Disease; Dose; DsRed; Effectiveness; Embryo; Endothelial Cells; Erythrocytes; Event; Fee-for-Service Plans; Fluorescence; Fluorine; Frequencies; Future; Glossary; Goals; Green Fluorescent Proteins; Hematopoietic stem cells; Histology; Image; Image Analysis; Imagery; Imaging technology; Implant; In Vitro; Injection of therapeutic agent; International; Intestines; Isotopes; Joint Ventures; Label; Laboratories; Licensing; Location; Magnetic Resonance Imaging; Magnetism; Manuals; Medicine; Methods; Molecular; Mus; Organ; Palpable; Parkinson Disease; Phase; Phenotype; Photons; Population; Positioning Attribute; Positron-Emission Tomography; Prevention; Probability; Proteins; Public Health; Reporter; Reporter Genes; Reporting; Research; Research Contracts; Research Personnel; Risk; Risk Estimate; Robotics; Scheme; Screening procedure; Smooth Muscle; Societies; Sorting - Cell Movement; Spinal cord injury; Staging; System; Tail; Techniques; Technology; Technology Assessment; Teratoma; Testing; Tetrachlorodibenzodioxin; Therapeutic; Time; Tissues; Undifferentiated; Universities; Veins; X-Ray Computed Tomography; base; cellular imaging; commercialization; cryostat; embryonic stem cell; fluorescence microscope; gamma Actin; genetically modified cells; imaging modality; improved; in vivo; instrumentation; molecular imaging; prevent; progenitor; promoter; public health relevance; red fluorescent protein; stem cell population; stem cell therapy; tomography; tool; urinary

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）：在这些细胞的广泛治疗应用成为可能之前，评估分化胚胎干细胞群体形成畸胎瘤的潜力的能力是迫切需要的。凯斯西储大学和BioInVision公司的研究人员将共同创建平台技术，以验证现有的胚胎干细胞分离和纯化范例（如抗体标记和荧光活化细胞分选），以及未来的纯化范例。生物平台由胚胎干细胞组成，当未分化时报告一个荧光标记（红色），当内皮谱系分化时报告另一个荧光标记（绿色）。分选将通过体外培养绿色细胞，在长期培养中培养这些细胞，并确定任何红色未分化细胞的外观来评估。为了评估体内细胞，我们将使用BioInVision独特的冷冻成像技术，该技术可以检测小鼠体内的单个荧光细胞。简而言之，低温成像系统包括一个大舞台，具有特殊成像功能的电动低温恒温器；一种改良的亮场/荧光显微镜；还有一个机器人xyz成像系统定位器，所有这些都是由控制系统完全自动化的，可以对整个鼠标进行切片和平铺成像。基本的实验模式将包括分类分化（绿色）细胞并通过尾部注射将其植入。几天后，冷冻成像将用于识别畸胎瘤和/或未分化（红色）细胞与形成畸胎瘤的潜力。冷冻成像平台比其他成像方法（如生物发光、MRI和PET）具有明显的优势，因为它不仅能够检测大的显性畸胎瘤，而且还能发现将来可能形成畸胎瘤的单细胞。