シンポジウム 3

Recent Progress in Human Epithelial CellCulture System and Its Application toCanter Research

Tohru Kiyono
Division of Carcinogenesis and Cancer Prevention
Department of Cell Culture TechnologyNational Cancer Center Research Institute, Japan

Unlike in a conventional culture system, most somatic epithelial stem cells show higher[Honors, Committees and Public Service]telomerase activity and proliferate virtually infinitely in vivo. Therefore, if the in vivo niche

1998

Sugiura Award of the Japanese Society for Virology

1999

Promoting Award of the Japanese Cancer Association

2010

Tamiya Award of Foundation for Promotion of Cancer Research

2017

Project Officer for eASIA.

for epithelial stem cells can be mimicked in vitro, it is possible to avoid cellular senescence.
Indeed, recent progress in culture system indicates that at least several human epithelial cells
can be infinitely propagated in culture. Liu et al. improved the culture system for keratinocyteswith 3T3 feeder layer by adding ROCK inhibitor, Y-27632. Sato and Clevers established

Editorial Board for Future Virology, Cancer Science, Japanese Journal of Clinical Oncology and American
Journal of Cancer Research

RSpondin-based 3D culture system in which LGR5-positive stem cells from multiple organs canform virtually ever-expanding epithelial organoids. These cells can be used as cells-of-origin[References]for various cancers to make in vitro carcinogenesis models of a given cancer. These culture

1. Ghani FI, Dendo K, Watanabe R, Yamada K, Yoshimatsu Y, Yugawa T, Nakahara T, Tanaka K,

methods can be also used for establishment of not only normal human cells but also cancer celllines. In this symposium, I would like to introduce recent advance of these culture methods

Yoshida H, Yoshida M, Ishikawa M, Goshima N, Kato T and Kiyono T.

An ex-vivo culture systemof ovarian cancer faithfully recapitulating the pathological features of primary tumors. Cells, 2019, inpress.

and its application to establish ovarian cancer cell lines. We developed a novel culture method

2. Inagawa, Y.; Yamada, K.; Yugawa, T.; Ohno, S.; Hiraoka, N.; Esaki, M.; Shibata, T.; Aoki, K.; Saya,

which can support proliferation of almost all primary epithelial ovarian cancer cells, as well asH.; Kiyono, T., A human cancer xenograft model utilizing normal pancreatic duct epithelial cells

primary normal human oviductal epithelial cells. Cancer cells from fresh or frozen specimens

conditionally transformed with defined oncogenes. Carcinogenesis 2014, 35: 1840-6.

were enriched by the anti-EpCAM antibody-conjugated magnetic beads, plated on Matrigelcoated plate and cultivated under the optimized culture conditions. Seventeen newly establishedovarian cancer cell lines, which included all four major histotypes of ovarian cancer, wereconfirmed to express histotype-specific markers in vitro. Some of the cell lines from all the fourhistotypes, except mucinous type, generated tumors in immune-deficient mice and the xenograft
tumor tissues recapitulated the corresponding original tissues faithfully. Furthermore, withpoorly tumorigenic cell lines including mucinous type, we developed a novel xenograft model
which could reconstruct the original tissue architecture through forced expression of a set ofoncogenes followed by its silencing. With combination of the novel culture method and cellderived xenograft system, virtually every epithelial ovarian cancer can be reconstituted in micein a timely fashion.

Tohru Kiyono, M.D., D.M.Sc
1984
Nagoya University School of Medicine, M.D.
1984-1996 Intern, Kyoritsu Hospital, Nagoya, Japan
1986-2002 Research Staff, Senior Research Staff and Section Head, Lab. of Viral
Oncology, Aichi Cancer Center, Research Institute, Nagoya, Japan
1996-1998 Fred Hutchinson Cancer Research Center, Washington, USA
2002
Chief, Division of Virology, National Cancer Center Research Institute2015
Chief, Division of Carcinogenesis and Cancer Prevention, National Cancer
Center Research Institute1415