CHARACTERIZE STRUCT & FUNCT OF HEPATOCYTE SPHEROIDS & USE IN TISSUE ENGINEERING

表征结构

• 批准号: 6117305
• 负责人: WEI-SHOU HU
• 金额: $ 1.13万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6117305
• 关键词: animal tissue; bioengineering /biomedical engineering; bioimaging /biomedical imaging; biomedical resource; enzymes; microscopy

Hepatocyte spheroids exhibit enhanced liver-specific function compared to hepatocytes cultured as a monolayer. Ultrastructural analysis shows that they also appear to form bile cananlicular-like structures, indicating a more tissue-like structure. Liver-specific cytochrome P450 activity, measured in situ using confocal. microscopy, increases over the course of spheroid self-assembly and decreases as cells disassemble into a monolayer. Examination of the diffusion of a fluorescent tracer confirms the formation of a ductal network within spheroids. In order to better demonstrate the formation of differentiated structure, more specific structural markers are being investigated. Preliminary studies have begun to examine the distribution of cytoskeleton and adhesioti receptors over the course of spheroid assembly. The ultimate goals are to understand how hepatocytes recognize each other to begin to form spheroids and when the transition to enhanced activity occurs in order to better exploit hepatocytes for use in tissue engineering applications, such as a bioartificial liver device. We will use the multiphoton imaging system at the EWR to study spheroids labeled by immunofluorescence. We anticipate that the deep-sectioning capabilities of the multiphoton system should allow us to see structures within the spheroids that are inaccessible by conventional confocal microscopy techniques.

肝细胞球体表现出增强的肝脏特异性功能 与单层培养的肝细胞相比。超微结构 分析表明，它们似乎也形成了胆小管样 结构，表明更像组织的结构。肝脏特异性 细胞色素P450活性，用共聚焦原位测定。 显微镜下，在球体自组装过程中增加和 随着细胞分解成单层而减少。考试的结果 荧光示踪剂的扩散证实了导管的形成 球体内的网络。为了更好地展示 形成差异化结构，结构更加具体化 正在对标志物进行调查。初步研究已经开始 检测细胞骨架和粘附素受体的分布。 椭球体组装的过程。最终的目标是理解 肝细胞如何相互识别开始形成球体和 当向增强活动过渡时，为了更好地 开发肝细胞用于组织工程应用，如 作为生物人工肝装置。我们将使用多光子成像技术 EWR研究免疫荧光标记球体的系统。 我们预计多光子的深切片能力 系统应该允许我们看到椭球体内的结构 传统的共聚焦显微镜技术无法获得。