Engineering of a sugar-derivatized porous network for hepatocyte culture.
Many tissue engineering applications require a scaffold or template conducive to cell attachment and maintenance of functions. It may also be advantageous in some cases for these scaffolds to have a controlled porous architecture to facilitate cellular or tissue ingrowth. In this study, we have engineered a porous carbohydrate-derivatized substrate for hepatocyte culture. Polystyrene foams, with pore sizes up to 100 microns, fabricated by phase separation from a homogeneous naphthalene solution, were derivatized with lactose and heparin, both of which are known to promote rat hepatocyte attachment and maintenance of its differentiated functions. Rat hepatocytes cultured on these derivatized foams exhibited a rounded cellular morphology with many microvilli evident on the surface of the cells. The hepatocytes showed an increase in albumin secretion for the first 3 days of culture in a defined, serum-free medium, and dropped back to initial levels by the end of 7 days. The production of cytochrome P450-dependent hydroxytestosterone metabolites were also measured. Two testosterone metabolites were maintained and five others were present but decreased over a culture period of 1 week. These carbohydrate-derivatized porous substrates may be useful for large-scale culture of hepatocytes, toxicology screening and for use in a liver assist device.
Gutsche, AT; Lo, H; Zurlo, J; Yager, J; Leong, KW
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