Probing Combinatorial Hepatocellular Microenvironments

描述（由申请方提供）：肝细胞中肝脏特异性功能的稳定性对于基于肝细胞的肝病治疗的成功至关重要。微环境因素如可溶性刺激物、细胞外基质（ECM）以及与邻近细胞的相互作用的影响被认为在肝细胞表型的调节中是重要的。由于历史上很难相互独立地调节这些微环境线索，因此尚不清楚它们如何合作对肝细胞功能产生影响。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，该平台利用机器人点样技术在组合微流体环境中在ECM微域上培养肝细胞。在具体目标1中，将设计和表征平台。在具体目标2中，该平台将用于探测肝细胞对HGF、TGF β和TGF α的组合的响应，同时铺在层粘连蛋白、胶原蛋白I和纤连蛋白上。可溶性和不溶性线索之间的相互作用的清晰图像将是未来设计和实施基于肝细胞的治疗肝病的基础。

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.