STRUCT & FUNCT OF HEPATOCYTE SPHEROID & USE IN TISSUE ENGINEERING

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 用共聚焦显微镜原位测定细胞色素P450活性， 在球体自组装过程中增加，并随着 细胞分解成单层。关于A的扩散的考察 荧光示踪剂证实在体内形成了导管网络 椭球体。为了更好地论证它的形成 分化的结构，更具体的结构标记正在被 调查过了。初步研究已经开始检查 细胞骨架和黏附受体在慢性阻塞性肺疾病过程中的分布 椭球体组件。最终目标是了解如何 肝细胞相互识别开始形成球体，以及何时 向增强活动的过渡发生在ORDA中，以更好地利用 用于组织工程应用的肝细胞，例如 生物人工肝装置。我们将使用多光子成像技术 IMR系统研究免疫荧光标记的球体。 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。

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 adhesion 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 orda 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 IMR 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.