A stem cell medium containing neural stimulating factor induces a pancreatic cancer stem-like cell-enriched population

Watanabe,Yusaku; Yoshimura,Kiyoshi; Yoshikawa,Koichi; Tsunedomi,Ryoichi; Shindo,Yoshitaro; Matsukuma,Sou; Maeda,Noriko; Kanekiyo,Shinsuke; Suzuki,Nobuaki; Kuramasu,Atsuo; Sonoda,Kouhei; Tamada,Koji; Kobayashi,Sei; Saya,Hideyuki; Hazama,Shoichi; Oka,Masaaki

doi:10.3892/ijo.2014.2603

A stem cell medium containing neural stimulating factor induces a pancreatic cancer stem-like cell-enriched population

Authors:
- Yusaku Watanabe
- Kiyoshi Yoshimura
- Koichi Yoshikawa
- Ryoichi Tsunedomi
- Yoshitaro Shindo
- Sou Matsukuma
- Noriko Maeda
- Shinsuke Kanekiyo
- Nobuaki Suzuki
- Atsuo Kuramasu
- Kouhei Sonoda
- Koji Tamada
- Sei Kobayashi
- Hideyuki Saya
- Shoichi Hazama
- Masaaki Oka
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Affiliations: Department of Digestive Surgery and Surgical Oncology, Yamaguchi University School of Medicine, Ube, Yamaguchi 755-8505, Japan, Department of Neurosurgery, Yamaguchi University School of Medicine, Ube, Yamaguchi 755-8505, Japan, Department of Molecular Pharmacology, Yamaguchi University School of Medicine, Ube, Yamaguchi 755-8505, Japan, Department of Ophthalmology, Yamaguchi University School of Medicine, Ube, Yamaguchi 755-8505, Japan, Department of Immunology and Cell Signaling Analysis, Yamaguchi University School of Medicine, Ube, Yamaguchi 755-8505, Japan, Department of Molecular Physiology and Medical Bioregulation, Yamaguchi University School of Medicine, Ube, Yamaguchi 755-8505, Japan, Division of Gene Regulation, Institute for Advanced Medical Research, Graduate School of Medicine, Keio University, Shinjuku-ku, Tokyo 160-8582, Japan
Published online on: August 14, 2014 https://doi.org/10.3892/ijo.2014.2603
Pages: 1857-1866
Copyright: © Watanabe et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

Cancer stem cells (CSCs) have been studied for their self-renewal capacity and pluripotency, as well as their resistance to anticancer therapy and their ability to metastasize to distant organs. CSCs are difficult to study because their population is quite low in tumor specimens. To overcome this problem, we established a culture method to induce a pancreatic cancer stem-like cell (P-CSLC)-enriched population from human pancreatic cancer cell lines. Human pancreatic cancer cell lines established at our department were cultured in CSC-inducing media containing epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), leukemia inhibitory factor (LIF), neural cell survivor factor-1 (NSF-1), and N-acetylcysteine. Sphere cells were obtained and then transferred to a laminin-coated dish and cultured for approximately two months. The surface markers, gene expression, aldehyde dehydrogenase (ALDH) activity, cell cycle, and tumorigenicity of these induced cells were examined for their stem cell-like characteristics. The population of these induced cells expanded within a few months. The ratio of CD24high, CD44high, epithelial specific antigen (ESA) high, and CD44variant (CD44v) high cells in the induced cells was greatly enriched. The induced cells stayed in the G0/G1 phase and demonstrated mesenchymal and stemness properties. The induced cells had high tumorigenic potential. Thus, we established a culture method to induce a P-CSLC-enriched population from human pancreatic cancer cell lines. The CSLC population was enriched approximately 100-fold with this method. Our culture method may contribute to the precise analysis of CSCs and thus support the establishment of CSC-targeting therapy.

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