Hepatocyte-based applications, such as metabolism/hepatotoxicity testing of drug like candidates in drug discovery, require optimal in vitro culture model for hepatocyte functional maintenance. Despite of the rapid emerging of novel hepatocyte 3D culture model with high fidelity of in vivo mimicry, Big Pharmas currently still prefer simple 2D culture model (i.e. 2D hepatocyte monolayer on natural extracellular matrix-coated microplate) and neglect the complex 3D culture models due to their difficulties to be adapted to the automated high-throughput screening platform. Conventional cell culture microplates coated with natural extracellular matrix allow hepatocytes to adhere tightly as two-dimensional (2D) monolayer, but these anchored hepatocytes rapidly lose their differentiated functions. In this thesis, we have developed a novel 3D hepatocyte monolayer culture to improve the current 2D hepatocyte monolayer culture, which can be readily applied for high throughput drug testing and potentially useful for other hepatocyte-based applications such as bioreactors or for cell maintenance in the bioartificial-liver assisted devices.

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