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Directed differentiation of pluripotent stem cells to functional hepatocytes.

Methods in molecular biology (Clifton, N.J.) (2013-04-03)
Philip Roelandt, Jolien Vanhove, Catherine Verfaillie
ABSTRACT

Differentiation of human stem cells to hepatocytes is crucial for industrial applications as well as to develop new therapeutic strategies for liver disease. The protocol described here, using sequentially growth factors known to play a role in liver embryonic development, efficiently differentiates human embryonic stem cells (hESC) as well as human-induced pluripotent stem cells (hiPSC) to hepatocytes by directing them through defined embryonic intermediates, namely, mesendoderm/definitive endoderm and hepatoblast and hepatocyte phenotype. After 28 days, the final differentiated progeny is a mixture of cells, comprising cells with characteristics of hepatoblasts and a smaller cell fraction with morphological and phenotypical features of mature hepatocytes. An extensive functional characterization of the stem cell progeny should be used to confirm that differentiated cells display functional characteristics of mature hepatocytes including albumin secretion, glycogen storage, and several detoxifying functions such as urea production, bilirubin conjugation, glutathione S-transferase activity, cytochrome activity and drug transporter activity.

MATERIALS
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Brand
Product Description

Sigma-Aldrich
MCDB 201 Medium, With trace elements, L-glutamine and 30 mM HEPES, powder, suitable for cell culture