There is a growing demand for the expansion and differentiation of stem cells for cell therapies, tissue engineering, and model systems for drug screening. Current methods for stem cell production are based on the use of batch tissue culture flasks, which have several drawbacks. In this paper we report on the use of a crossed hollow fiber membrane bioreactor for the expansion and differentiation of embryonic liver cells, which have been used as an alternative model of human liver progenitor cells. The bioreactor is based on two bundles of fiber (PEEK-WC HF and PES-HE) which are cross-assembled in an alternating manner. This bioreactor geometry ensures high mass exchange of nutrients and metabolites, which is important for cell proliferation and differentiation. The membrane bioreactor, thanks to its optimized fluid dynamics and mass transport and the adequate surface properties of hollow fibers, was able to guide the expansion and differentiation of liver progenitor cells in mature hepatocytes as demonstrated by their expression of liver specific functions in terms of urea synthesis, albumin production, and diazepam drug biotransformation.