Probing Combinatorial Hepatocellular Microenvironments

描述(由申请人提供)：肝细胞中肝脏特异性功能的稳定性是以肝细胞为基础的肝病治疗成功的关键。微环境因素，如可溶性刺激、细胞外基质(ECM)以及与邻近细胞的相互作用，在调节肝细胞表型方面被认为是重要的。由于历史上很难相互独立地调节这些微环境信号，它们如何合作对肝细胞功能施加影响仍不清楚。为了系统地研究微环境线索的相互作用，提出了一种新的平台，利用机器人斑点技术在组合微流控环境中的ECM微域上培养肝细胞。在具体目标1中，将对平台进行设计和表征。JN特定目标2，该平台将用于检测肝细胞对HGF、EOF和TGFα组合的反应，同时将其电镀在层粘连蛋白、I型胶原和纤维连接蛋白上。清楚地了解可溶和不可溶线索之间的相互作用将是未来设计和实施基于肝细胞的疗法治疗肝病的基础。

DESCRIPTION (provided by applicant): Stability of liver-specific functions in hepatocytes is critical to the success of hepatocyte-based therapies for liver disease. The influences of microenvironmental factors such as soluble stimuli, extracellular matrix (ECM), and interactions with neighboring cells are thought to be important in regulation of the hepatocyte phenotype. Because it has historically been difficult to modulate these micro-environmental cues independently from each other, it remains unclear how they cooperate to exert their influence on hepatocellular function. To systematically investigate the interaction of micro-environmental cues, a novel platform is proposed that exploits robotic spotting technology to culture hepatocytes atop ECM microdomains within combinatorial microfluidic environments. In specific aim 1, the platform will be designed and characterized. Jn specific aim 2, the platform will be used to probe hepatocellular responses to combinations of HGF, EOF, and TGFalpha while plated upon laminin, collagen I, and fibronectin. A clear picture of the interplay between soluble and insoluble cues will be fundamental to the future design and implementation of hepatocyte-based therapies to treat liver disease.