Faculty Profiles - SAKAGUCHI Masakiyo

MISC - SAKAGUCHI Masakiyo

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Adenovirus-mediated p21 gene transfer enhances differentiated hepatic phenotypes of immortalized human hepatocytes.

N Kobayashi, H Noguchi, T Totsugawa, T Watanabe, T Matsumura, M Maruyama, T Matsumoto, T Fujimara, N Tanaka, M Sakaguchi

HEPATOLOGY 34 ( 4 ) 305A - 305A 2001.10

Expansion of human hepatocyte populations by a retroviral gene transfer of simian virus 40 large T antigen

N Kobayashi, KA Westerman, T Taguchi, M Sakaguchi, T Fujiwara, H Urata, N Kishimoto, N Hayashi, S Nakaji, T Murakami, P Leboulch, N Tanaka

ASAIO JOURNAL 47 ( 5 ) 481 - 485 2001.9

Language：English Publisher：LIPPINCOTT WILLIAMS & WILKINS

A hybrid artificial liver (HAL) could be used to treat acute liver failure or to serve as a temporary support until orthotopic liver transplantation is available. Primary human hepatocytes are ideal as a source of hepatic function in a HAL device. However, the worldwide shortage of human livers available for hepatocyte isolation severely limits this form of therapy. A possible alternative is to use a tightly regulated cell line that can be economically grown in culture to have differentiated liver function. In this work, human hepatocytes were immortalized with a retroviral vector SSR#69 expressing the genes of simian virus 40 large T antigen and herpes simplex virus-thymidine kinase. One of the resulting clones, NKNT-3, showed the gene expression of differentiated liver function and were sensitive to the antiviral agent ganciclovir. When transplanted into the spleen of rats subjected to 90% hepatectomy, NKNT-3 cells prolonged the survival of 90% hepatectomized rats. The cells provide the advantages of unlimited availability, sterility, uniformity, and freedom from pathogens. This work represents a potential novel strategy for resolving the organ shortage that currently limits the use of primary human hepatocytes to develop a HAL.

DOI： 10.1097/00002480-200109000-00017

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Construction of a differentiated human hepatocyte cell line expressing the herpes simplex virus-thymidine kinase gene

N Kobayashi, M Miyazaki, KA Westerman, H Noguchi, M Sakaguchi, T Totsugawa, T Watanabe, T Matsumura, T Fujiwara, P Leboulch, N Tanaka, M Namba

ASAIO JOURNAL 47 ( 5 ) 476 - 480 2001.9

Language：English Publisher：LIPPINCOTT WILLIAMS & WILKINS

Transient support using a hybrid artificial liver (HAL) device is a promising treatment for the patients with acute liver failure. Primary human hepatocytes are an ideal source for HAL therapy; however, the number of human livers available for hepatocyte isolation is limited by competition for use in whole organ transplantation. To overcome this problem, we previously established a highly differentiated human fetal hepatocyte cell line OUMS-29. Considering the potential risk when using these genetically engineered cells in humans, additional safeguards should be added to make the cells more clinically useful. In this work, the herpes simplex virus thymidine kinase (HSVtk) gene was retrovirally introduced into OUMS-29 cells. One of the HSVtk-expressed clones, OUMS-29/thymidine kinase (TK), grew in chemically defined serum free medium and expressed the genes of albumin, asialoglycoprotein receptor, glutamine synthetase, glutathione-S-transferase pi, and blood coagulation factor X. In vitro sensitivity of the cells to ganciclovir was evaluated. Intrasplenic transplantation of 50 X 10(6) OUMS-29/TK cells prolonged the survival of 90% hepatectomized rats compared with medium injection alone (control). In the present study, we have established highly differentiated immortalized human hepatocytes with tight regulation. The cells may be clinically useful for HAL treatment.

DOI： 10.1097/00002480-200109000-00016

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S100Cタンパク質のアクチンとの結合及びその意義

阪口政清, 宮崎正博, 難波正義, 許南浩

Tissue culture research communications : the journal of experimental & applied cell culture research = 組織培養研究 20 ( 2 ) 108 - 108 2001.7

レンチウイルスベクターによるヒト血管内皮細胞への効果的な遺伝子導入.

都津川敏範, 小林直哉, 丸山昌伸, 興津輝, 野口洋文, 松村年久, 渡辺剛正, 阪口政清, 藤原俊義, 田中紀章

Organ Biology 2001

ヒト線維芽細胞の不死化, 癌化にともなう細胞内タンパク質の変化 : 酸化還元系酵素の発現の変化について

近藤格, 阪口政清, 難波正義

生物物理化学 = Journal of Electrophoresis 44 35 - 35 2000.10

Effects of Oxygen Concentrations on Human Fibroblasts Treated with Fe^<3+>-NTA

PU Hong, SAKAGUCHI Masakiyo, KONDO Tadashi, KONDO Asami, JIANG H-X., KAWABATA Teruyuki, NAMBA Masayoshi

19 ( 2 ) 78 - 78 2000.6

Relationship between Contact Inhibition and Intranuclear S100C of Normal Human Fibroblasts

SAKAGUCHI Masakiyo, MIYAZAKI Masahiro, KOUCHI Hirosuke, KONDO Tadashi, NAMBA Masayoshi

19 ( 2 ) 87 - 87 2000.6

【臓器移植と医工学その現況と展望】急性肝不全に対する不死化ヒト肝細胞の移植

小林直哉, 野口洋文, 田中紀章, 深谷憲一, 阪口政清, 井上祐介, 宮崎正博, 難波正義

今日の移植 13 ( 3 ) 216 - 221 2000.5

SF10-3 不死化ヒト臍帯静脈内皮細胞株(HNKT-1)の樹立

野口洋文, 小林直哉, 宮崎正博, 井上祐介, 阪口政清, 藤原俊義, 田中紀章

日本外科学会雑誌 101 129 - 129 2000.3

ペプシンを産生するラット胃粘膜上皮細胞株(OUMS-37)の樹立と分化誘導の検討

濮紅, 湯浅貴恵, 近藤麻美, 阪口政清, 高崇, 稲田憲一, 難波正義

Tissue culture research communications : the journal of experimental & applied cell culture research = 組織培養研究 18 ( 1 ) 63 - 63 1999.3

細胞外ATPは不死化ヒト線維芽細胞の増殖をヒト正常線維芽細胞に比べより強く抑制する

井上裕介, 阪口政清, 難波正義

Tissue culture research communications : the journal of experimental & applied cell culture research = 組織培養研究 18 ( 1 ) 67 - 67 1999.3

P-986 ラット急性肝不全モデルにおける不死化胎児肝細胞OUMS-29の脾臓内移植の効果

小林直哉, 宮崎正博, 井上裕介, 阪口政清, 近藤麻美, 田中紀章, 難波正義

日本外科学会雑誌 100 558 - 558 1999.2

ATP inhibition of proliferation of immortalized human fibroblasts is greater than that of normal human diploid fibroblasts

B Katayama, M Sakaguchi, JW Li, H Pu, Y Inoue, M Namba

INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE 2 ( 5 ) 603 - 606 1998.11

Language：English Publisher：PROFESSOR D A SPANDIDOS

It is known that cancers develop by a multi-step process. Normal cells are first immortalized, and then transformed into tumorigenic cells. Normal human cells are very rarely immortalized, but once they are, they are relatively easily transformed into tumorigenic cells. This indicates that the immortalization step plays a critical part in the development of human cancers. Thus, elucidation of the mechanisms of this step would shed light on the process of carcinogenesis in human cells. To understand the causes of immortalization, it is important to determine the differences in cellular phenotype between immortalized and normal human cells. In this study, we found that immortalized human fibroblasts were more sensitive to the growth inhibitory effects of ATP than normal human fibroblasts. ADP was as effective as ATP, but AMP, adenosine, and phosphoric acid were not. These results indicate that a high-energy bound of ATP and ADP may contribute to the growth inhibition of the cells. When the immortalized cells were pulse-labeled with [P-32]-ATP, 30-, 31-, 33- and 40-kDa membrane fraction proteins were more prominently labeled in the immortalized cells than in the normal cells. At present, the characteristics of these proteins are being investigated.

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Differential intracellular localization of transferrin produced in human fibroblasts and that incorporated from culture medium

SAKAGUCHI M., KONDO T., NAMBA M.

17 ( 1 ) 46 - 46 1998.3

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