Miki Atsushi, Narushima Michiki, Okitsu Teru, Takeno Yuichi, Soto-Gutierrez Alejandro, Rivas-Carrillo Jorge David, Navarro-Alvarez Nalú, Chen Yong, Tanaka Kimiaki, Noguchi Hirofumi, Matsumoto Shinichi, Kohara Michinori, Lakey Jonathan R T, Kobayashi Eiji, Tanaka Noriaki, Kobayashi Naoya
Department of Surgery, Okayama University Graduate School of Medicine and Dentistry, 2-5-1 Shikata-cho, Okayama 700-8558, Japan.
Department of Transplant Surgery, Kyoto University Hospital, 54 Seigoin-Kawaracho, Sakyo-ku, Kyoto 606-8507, Japan.
Cell Transplant. 2006 Apr;15(4):325-334. doi: 10.3727/000000006783981882.
Development of an efficient preculture system of islets is ideal. Toward that goal, we constructed a human pancreatic islet-derived fibroblast cell line MNNK-1 for a source as a coculture system for freshly isolated islets to maintain islet functions. Human pancreatic islet cells were nucleofected with a plasmid vector pYK-1 expressing simian virus 40 large T antigen gene (SV40T) and hygromycin resistance gene (HygroR). One of the transduced cell lines, MNNK-1, was established and served as a feeder cell in the coculture for freshly isolated mouse, rat, and pig islets. Morphology, viability, and glucose-responding insulin secretion were analyzed in the coculture system. MNNK-1 cells were morphologically spindle shaped and were negative for pancreatic endocrine markers. MNNK-1 cells were positive for α-smooth muscle actin and collagen type I and produced fibroblast growth factor. Coculture of the mouse, rat, and pig islets with MNNK-1 cells maintained their viability and insulin secretion with glucose responsiveness. A human pancreatic islet-derived fibroblast cell line MNNK-1 was established. MNNK-1 cells were a useful means for maintaining morphology and insulin secretion of islets in the coculture system.
