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）：评估分化胚胎干细胞种群的畸胎瘤形成潜力的能力代表了在这些细胞的广泛治疗应用之前的迫切需求。 Case Western Reserve University和Bioinvision，Inc。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式（例如抗体标记和荧光激活的细胞分选）以及未来的纯化范式。该生物平台由胚胎干细胞组成，这些干细胞在未分化时报告了一个荧光标记（红色），而在区分内皮谱系时进行了另一个标记（绿色）。分类将在体外通过镀绿细胞，在长期培养中种植这些细胞以及确定红色未分化细胞的任何外观来评估排序。为了评估体内细胞，我们将使用生物引导的独特的冷冻成像技术，该技术可以检测小鼠中的单个荧光细胞。简而言之，冷冻成像系统由一个大型舞台，机动的低温恒温器组成，具有特殊的成像功能。改良的，明亮的场/荧光显微镜；以及机器人XYZ成像系统定位器，所有这些系统都是由控制系统完全自动化的，以允许整个鼠标的切片和瓷砖成像。基本的实验范式将包括对分化（绿色）细胞进行分类并通过尾注射植入它们。几天后，冷冻成像将用于鉴定畸胎瘤和/或未分化的（红色）细胞，具有形成畸胎瘤的潜力。与其他成像方法（例如生物发光，MRI和PET）相比，冷冻成像平台代表了一个明显的优势，因为它不仅能够检测出大型明显的畸胎瘤，而且还可以找到可能在未来形成TeraTomas的单个细胞。 公共卫生相关性：该提案与公共卫生的问题非常关注，因为它的长期目标是提供一种评估胚胎干细胞选择和纯化策略的有效性的方法，然后再进行治疗。畸胎瘤形成的能力是阻止临床上使用胚胎干细胞的主要障碍，例如糖尿病和帕金森氏病和脊髓损伤。