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 subst...

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Bibliographic Details
Published in:Journal of biomaterials science. Polymer ed. 2000-01, Vol.11 (7), p.731-745
Main Authors: Fujii, Yasuo, Nakazawa, Kohji, Funatsu, Kazumori
Format: Article
Language:English
Subjects:
Ammonia - analysis
Animals
Biocompatible Materials
Cell Culture Techniques - methods
Cell Nucleus - ultrastructure
Cell Size
Cells, Cultured
Chromatography, Gel
CONDITIONED
Culture Media, Conditioned - analysis
FOAM
HEPATOCYTE
Hepatocytes - cytology
Hepatocytes - physiology
Liver, Artificial
Male
MEDIUM
POLYURETHANE
Polyurethanes
PRIMARY
Rats
Rats, Wistar
Serum Albumin - analysis
SPHEROID
Ultrafiltration
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Summary: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.
ISSN:0920-5063
1568-5624
DOI:10.1163/156856200743986