Maintenance of mouse, rat, and pig pancreatic islet functions by coculture with human islet-derived fibroblasts.
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.
Miki
Atsushi
A
Department of Surgery, Okayama University Graduate School of Medicine and Dentistry, 2-5-1 Shikata-cho, Okayama 700-8558, Japan.
Narushima
Michiki
M
Okitsu
Teru
T
Takeno
Yuichi
Y
Soto-Gutierrez
Alejandro
A
Rivas-Carrillo
Jorge David
JD
Navarro-Alvarez
Nalú
N
Chen
Yong
Y
Tanaka
Kimiaki
K
Noguchi
Hirofumi
H
Matsumoto
Shinichi
S
Kohara
Michinori
M
Lakey
Jonathan R T
JR
Kobayashi
Eiji
E
Tanaka
Noriaki
N
Kobayashi
Naoya
N
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
D000199:Actins; D024042:Collagen Type I; D007328:Insulin; D005947:Glucose; D015774:Ganciclovir