STEM CELL TRACKING: IN VITRO AND IN VIVO APPLICATIONS

干细胞追踪：体外和体内应用

• 批准号: 7358296
• 负责人: EDWARD W HSU
• 金额: $ 1.03万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7358296
• 关键词: STEM; CELL; TRACKING; VITRO; VIVO

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. To track labeled stem/progenitor adherent cells as they migrate and seed thoughout in-vivo tissue segements. The goals are: - To help determine the best seeding techniques (e.g. either within tissues or via arterial/venus injections) that would allow most adherent stem cells to remain at the injured tissue sites. - To develop unique cell monitoring capabilities, using PET, of in-vivo stem cells. In this process stem cells are pre-labeled with a thimadine kinase (TK) constructs using lenti vector technologies. After cell labeling, the inherent TK cleaves an FHBG PET probe to activate PET signaling. Here, long term investiagations of in-vivo cell migration and aggregation may be analyzed by allowing pre-labeled TK stemcells to adhere inside tissue components over nonspecific time fames - then activate PET signaling prior to image acquisition. - Using negative contrast MRI, we will concurrently (along with PET) monitor in-vivo stem cells as they are seeded within tissues sites. For this imaging modality, unique micro iron particles that include either A)outer dextran coatings, or B)specific receptor-ligand communications (e.g. EpCAM), are used to promote endocytosis of iron particles such that distinctive iron concentrations are located within the cells cytoplasm. This labeling proces is used to disrupt MR signaling and promote negative signal imaging. - To quantify the amount of cells needed to induce tissue repair. - To visualize a stem cell signal of the smallest aggregate size (hopefully single cell suspensions) as they seed themselves inside capillary beds of injured tissues. At present, stem cell therapies assume that seeded cells remain within the tissues of interest. In reality, many cells flow through the damaged tissues and implant themselves inside various capillary beds throughout the body. Improved tacking methods are necessary to visualize the dispersion of cells, to visualize cell aggregate sizes, and to specifically quantify stem cell numbers that remain at the tissue injury sites.

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。追踪标记的干/祖细胞贴壁细胞在体内组织片段中的迁移和接种。目标是：- 帮助确定最佳的接种技术（例如，在组织内或通过动脉/静脉注射），这将允许大多数粘附的干细胞留在受伤的组织部位。- 开发独特的细胞监测能力，使用PET，体内干细胞。在该过程中，使用慢病毒载体技术用噻吗啶激酶（TK）构建体预标记干细胞。细胞标记后，固有TK切割FHBG PET探针以激活PET信号传导。在此，体内细胞迁移和聚集的长期研究可以通过允许预标记的TK干细胞在非特异性时间范围内粘附在组织组分内来分析，然后在图像采集之前激活PET信号传导。- 使用负对比MRI，我们将同时（沿着PET）监测体内干细胞，因为它们被接种在组织部位内。对于这种成像模式，使用包括A）外葡聚糖涂层或B）特异性受体-配体通信（例如EpCAM）的独特的微铁颗粒来促进铁颗粒的内吞作用，使得独特的铁浓度位于细胞质内。该标记过程用于破坏MR信号传导并促进负信号成像。 - 以量化诱导组织修复所需的细胞数量。- 可视化最小聚集尺寸的干细胞信号（希望是单细胞悬浮液），因为它们在受损组织的毛细血管床内播种。目前，干细胞疗法假设接种的细胞保留在感兴趣的组织内。事实上，许多细胞流经受损组织，并将自己植入全身各种毛细血管床内。需要改进的定位方法来可视化细胞的分散，可视化细胞聚集体的大小，并具体量化保留在组织损伤部位的干细胞数量。