Maintenance of mouse, rat, and pig pancreatic islet functions by coculture with human islet-derived fibroblasts

Atsushi Miki, Michiki Narushima, Teru Okitsu, Yuichi Takeno, Alejandro Soto-Gutierrez, Jorge David Rivas-Carrillo, Nalú Navarro-Alvarez, Yong Chen, Kimiaki Tanaka, Hirofumi Noguchi, Shinichi Matsumoto, Michinori Kohara, Jonathan R.T. Lakey, Eiji Kobayashi, Noriaki Tanaka, Naoya Kobayashi

Research output: Contribution to journalArticle

27 Scopus citations

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 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.

Original languageEnglish (US)
Pages (from-to)325-334
Number of pages10
JournalCell Transplantation
Volume15
Issue number4
DOIs
Publication statusPublished - Aug 8 2006

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Keywords

  • Coculture
  • Islets
  • Simian virus 40 large T antigen

ASJC Scopus subject areas

  • Biomedical Engineering
  • Cell Biology
  • Transplantation

Cite this

Miki, A., Narushima, M., Okitsu, T., Takeno, Y., Soto-Gutierrez, A., Rivas-Carrillo, J. D., ... Kobayashi, N. (2006). Maintenance of mouse, rat, and pig pancreatic islet functions by coculture with human islet-derived fibroblasts. Cell Transplantation, 15(4), 325-334. https://doi.org/10.3727/000000006783981882