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Development of Highly Functional Long-Term Culture Method of Liver Slice Embedded in Agarose Gel for Bioartificial Liver

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It is difficult to a produce highly functional bioartificial liver (BAL) using only hepatocytes, because it is believed that liver-specific three-dimensional structure is necessary to maintain high function for BAL. But it is difficult to construct a culture system with liver-specific three-dimensional structure in vitro. To realize a highly functional culture system with liver-specific three-dimensional structure, we developed a culture system using liver slices that keep liver-specific architecture, such as liver lobule and hepatic microvascular system. Liver slices were embedded in agarose gel to maintain them under a moist and three-dimensional environment. We examined the viability and function of liver slices by using various shapes of agarose gel. Liver slices were cultured 1) under stationary condition (control), 2) directly embedded in gel, and 3) embedded in cylindrical gel for good drainage of medium and ventilation of air. The viability and function of the incubated liver slices were evaluated by LDH leakage, histomorphology, and immunohistochemistry. At 10 days, the morphological condition and function of liver slices embedded in cylindrical gel were maintained better than liver slices directly embedded in gel or in the stationary condition. We suggest that high functionality and morphological condition of liver slices could be maintained by embedding in cylindrical gel. In the future, it is possible that this method could be used to develop a highly functional bioartificial liver.
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Keywords: Agarose gel; Bioartificial liver; Liver slice; Three-dimensional culture

Document Type: Research Article

Affiliations: 1: *Department of Functional Polymer Science, Faculty of Textile Science and Technology, Shinshu University 3-15-1 Tokida, Ueda 386-8567, Japan 2: †Development of Organ Regeneration Institute of Organ Transplants, Reconstructive Medicine and Tissue Engineering, Shinshu University Graduate School of Medicine, 3-1-1 Asahi, Matsumoto 390-8621, Japan 3: ‡Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Director Research Center for Experimental Biology, Tokyo Institute of Technology, 4259 Nagatsuta-Cho, Midori-Ku, Yokohama-Shi 226-8501, Japan

Publication date: 2003-01-01

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