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KMID : 1094719980030020096
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Biotechnology and Bioprocess Engineering
1998 Volume.3 No. 2 p.96 ~ p.102
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Development of hepatocyte spheroids immobilization technique using alternative encapsulation method
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Kim Sung-Po
Lee Doo-Hoon
Park Jung-Keug
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Abstract
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Primary hepatocytes of small animals such as rat and rabbit were often used for the study of extracorporeal liver support systems. Freshly isolated rat hepatocytes form spheroids within two days when cultivated as suspension in spinner vessels. These spheroids showed enhanced liver specific functions and more differentiated morphology compared to hepatocytes cultured as monolayers. However, shear stress caused by continuous agitation deteriorated spheroids gradually. In this work we immobilized spheroids to prolong liver specific activities. First, hepatocyte spheroids were suspended in collagen solution containing calcium chloride and then dropped into alginate solution. A thin layer of calcium alginate was formed around the droplet and then was removed after the inner collagen was gelled by treatment of sodium citrate buffer. Spheroids embedded in collagen-gel bead maintained liver specific functions such as albumin secretion rate longer than hepatocyte spheroids exposed to shear stress. Therefore, we suggest that this immobilization technique may offer an effective long-term hepatocyte cultivation and facilitate the development of a bioartificial liver support device.
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KEYWORD
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Bioartificial liver, hepatocyte spheroid, collagen, encapsulation
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