Induction and High Density Culture of Human Hepatoblasts from Fetal Hepatocytes with Suppressing Transformation(Molecular and Cell Biology)
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概要
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It is well known that it would be important to cultivate human hepatocytes of about 10^<10> cells at a high cell density, about 1×10^7 cells/cm^3, in the bioreactor for the development of bioartificial liver. However, since primary human hepatocytes lack an ability to proliferate in vitro, it is essential to establish a culture method for the proliferation of normal human hepatic stem cells as a cell source. In this study, it was found that human hepatoblasts, a kind of hepatic stem cells, were induced from human fetal hepatocytes while keeping the ability of proliferation by the treatment of 1mM sodium butyrate (SB) for 12d of culture. The transformation of hepatoblasts was evaluated by abnormal prothrombin (PIVKA-II) assay, which is a clinical marker for hepatocellular carcinoma. The PIVKA-II production rate of the cells was suppressed to the normal level under 1mM SB. The cells including hepatoblasts under 1mM SB attached to the porous hydroxyapatite carriers and proliferated to a high cell density of about 1×10^7 cells/cm^3 in the carriers. The liver-specific function, cytochrome P450 3A4 activity (4.2 pmol/mg protein/min) of the cells in the carriers under 1mM SB was comparable to that of primary human hepatocytes. Ammonia metabolizing activity (0.21μmol/10^6 cells/h) of the cells was also comparable to that of porcine hepatocytes used in the bioartificial liver. The PIVKA-II production rate of the cells in the carrier was suppressed to the normal level. These results suggested that induction of human hepatoblasts from fetal hepatocytes by the treatment of 1mM SB and proliferation of the cells at a high cell density using hydroxyapatite carriers should be one of the more promising culture methods for bioartificial liver developments.
- 2007-12-01
著者
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UEOKA Ryuichi
Division of Applied Chemistry, Graduate School of Kumamoto Institute of Technology
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Matsushita Taku
Division Of Applied Life Science Graduate School Of Engineering Sojo University
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Matsushita Taku
Sojo University
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Matsushita Taku
Graduate Course Of Applied Chemistry Sojo University
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Ueoka Ryuichi
Division Of Applied Life Science Graduate School Of Engineering Sojo University
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KIYOTA Akifumi
Division of Applied Life Science, Graduate School of Engineering, Sojo University
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Kiyota Akifumi
Division Of Applied Life Science Graduate School Of Engineering Sojo University
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