Intensive promotion of spheroid formation by soluble factors in a hepatocyte-conditioned medium
We developed a hybrid artificial liver and a drug metabolism simulator using polyurethane foam (PUF) in which primary hepatocytes spontaneously form functional spheroids. Gel filtration liquid chromatography
analysis of a hepatocyte-conditioned medium during spheroid formation showed that some substances secreted by primary rat hepatocytes accumulated advantageously inside the pores of PUF compared with outside.
Similar substances were detected in a hepatocyte-conditioned medium from a positively-charged surface by concentrating the substances using an ultrafiltration membrane of a molecular weight-cutoff of 50
kD. These substances were shown to act as soluble factors on freshly isolated primary rat hepatocytes to promote spontaneous and rapid spheroid formation, depending on their concentration by preventing
them from initially attaching and spreading on a positively-charged surface. In particular, using 50-fold concentrated substances, about 80% of total hepatocytes formed the floating spheroids within 72
h of culture. The resulting spheroids had a diameter distribution mainly ranging from 40 to 70 μm and expressed high-level liver-specific functions compared with a conventional monolayer.
Keywords: CONDITIONED; FOAM; HEPATOCYTE; MEDIUM; POLYURETHANE; PRIMARY; SPHEROID
Document Type: Research Article
Affiliations: Department of Chemical Systems and Engineering, Graduate School of Engineering, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka, 812-8581, Japan
Publication date: 30 August 2000
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