Hepatocyte-based applications, such as metabolism/hepatotoxicity drug testing for drug discovery and bioartificial-liver assisted devices are challenged by the lack of optimal in vitro culture model for hepatocyte functional maintenance and shortage of sustainable cell source. In this talk, two examples will be introduced to take advantage of 3D engineering configurations to tackle these challenges. In the first part, a novel 3D hepatocyte monolayer culture configuration was developed in a bioactive RGD/Galactose hybrid substrate to improve the gold-standard 2D hepatocyte monolayer culture, which can be readily applied for multi-well plate-based high throughput drug testing. In the second part, directed differentiation of human embryonic stem cells (hESCs) are achieved within a micro-patterned platform to recapitulate the embryonic liver development. Compared with 2D planar differentiation, the 3D micro-engineered platform generated more homogenous and mature hepatocyte-like cells with improved hepatic functions. These two 3D configurations for hepatic maintenance and differentiation represent significant step towards better control of hepatic cell fate for their wider applications in drug discovery and medical treatment for liver failures.