title: Maintenance of mouse, rat, and pig pancreatic islet functions by coculture with human islet-derived fibroblasts.
abstract: 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 alpha-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.
authors.affiliation: Department of Surgery, Okayama University Graduate School of Medicine and Dentistry, 2-5-1 Shikata-cho, Okayama 700-8558, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Miki
authors.forename: Michiki
authors.initials: M
authors.lastname: Narushima
authors.forename: Teru
authors.initials: T
authors.lastname: Okitsu
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Takeno
authors.forename: Alejandro
authors.initials: A
authors.lastname: Soto-Gutierrez
authors.forename: Jorge David
authors.initials: JD
authors.lastname: Rivas-Carrillo
authors.forename: Nalú
authors.initials: N
authors.lastname: Navarro-Alvarez
authors.forename: Yong
authors.initials: Y
authors.lastname: Chen
authors.forename: Kimiaki
authors.initials: K
authors.lastname: Tanaka
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Noguchi
authors.forename: Shinichi
authors.initials: S
authors.lastname: Matsumoto
authors.forename: Michinori
authors.initials: M
authors.lastname: Kohara
authors.forename: Jonathan R T
authors.initials: JR
authors.lastname: Lakey
authors.forename: Eiji
authors.initials: E
authors.lastname: Kobayashi
authors.forename: Noriaki
authors.initials: N
authors.lastname: Tanaka
authors.forename: Naoya
authors.initials: N
authors.lastname: Kobayashi
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D018920:Coculture Techniques / Q000379:methods; D024042:Collagen Type I / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism / Q000502:physiology; D015774:Ganciclovir / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism; D006801:Humans; D007328:Insulin / Q000378:metabolism; D007515:Islets of Langerhans / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D016513:Mice, SCID; D051381:Rats; D013552:Swine; D014022:Tissue Survival / Q000502:physiology
chemical_list: D000199:Actins; D024042:Collagen Type I; D007328:Insulin; D005947:Glucose; D015774:Ganciclovir