Galactosylated Poly(vinylidene difluoride) Hollow Fiber Bioreactor for Hepatocyte Culture
| Autor: | Hai-Quan Mao, Hong-Fang Lu, Hanry Yu, Ser-Mien Chia, Wei Seng Lim, Peng-Chi Zhang, Kam W. Leong |
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| Rok vydání: | 2005 |
| Předmět: |
Male
Cell Culture Techniques Serum albumin law.invention Bioreactors law Animals Fiber Rats Wistar Cells Cultured Tissue Engineering biology Chemistry General Engineering Bioartificial liver device Galactose Substrate (chemistry) Membranes Artificial Poloxamer Liver Artificial Rats Membrane Biochemistry Chemical engineering Transmission electron microscopy Hepatocytes biology.protein Polyvinyls Protein adsorption |
| Zdroj: | Tissue Engineering. 11:1667-1677 |
| ISSN: | 1557-8690 1076-3279 |
| DOI: | 10.1089/ten.2005.11.1667 |
| Popis: | To overcome the limitations of long-term expression of highly differentiated hepatocyte functions, we have developed a novel bioreactor in which hepatocytes are seeded in a ligand-immobilized hollow fiber cartridge. Galactosylated Pluronic polymer is immobilized on poly(vinylidene difluoride) (PVDF) hollow fiber surface through an adsorption scheme yielding a substrate with hepatocyte-specific ligand and a hydrophilic surface layer, which can resist nonspecific protein adsorption and facilitate cell binding to the galactose ligand. Interestingly, the galactosylated PVDF hollow fiber shows enhanced serum albumin diffusion across the membrane. Freshly isolated rat hepatocytes were seeded and cultured in the extralumenal space of the hollow fiber cartridge for 18 days in a continuously circulated system. Albumin secretion function of the seeded hepatocytes was monitored by analyzing circulating medium by enzyme-linked immunosorbent assay. Urea synthesis and P-450 function (7-ethoxycoumarin dealkylase activity) were measured periodically by doping the circulating medium with NH4Cl and 7-ethoxycoumarin, respectively. Hepatocytes cultured on galactosylated PVDF hollow fibers maintained better albumin secretion and P-450 functions than on unmodified and serum-coated PVDF hollow fibers when cultured in serum-containing medium. Morphological examination by scanning electron microscopy showed that hepatocytes cultured on galactosylated PVDF hollow fibers developed significant aggregation, in contrast to those cultured on unmodified PVDF fibers or on serum-coated PVDF fibers. Transmission electron microscopy images revealed that tight junctions and canaliculus-like structures formed in these aggregates. These results suggest the potential application of this galactosylated PVDF hollow fiber cartridge for the design of a bioartificial liver assist device. |
| Databáze: | OpenAIRE |
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