Establishment of a pancreatic stem cell line from fibroblast-derived induced pluripotent stem cells

BioMedical Engineering OnLine

Takashi Kuise⁵  Hirofumi Noguchi²  Hiroshi Tazawa⁵  Takashi Kawai⁵  Masaya Iwamuro³  Issei Saitoh⁶  Hitomi Usui Kataoka⁷  Masami Watanabe⁴  Yasufumi Noguchi¹  Toshiyoshi Fujiwara⁵ 

[1] Department of Socio-environmental Design, Hiroshima International University, Hiroshima 737-0112, Japan

[2] Department of Surgery, Chiba-East National Hospital, National Hospital Organization, Chiba 260-8712, Japan

[3] Department of Gastroenterology and Hepatology, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama 700-8558, Japan

[4] Department of Urology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8558, Japan

[5] Department of Gastroenterological Surgery, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8558, Japan

[6] Department of Pediatric Dentistry, Niigata University Graduate School of Medical and Dental Sciences, Niigata 951-8514, Japan

[7] Department of Primary Care and Medical Education, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama 700-8558, Japan

关键词: iPS cells;    Diabetes;    Mouse pancreatic stem cells;   

Others  :  793300
DOI  :  10.1186/1475-925X-13-64

 received in 2014-04-02, accepted in 2014-05-23,  发布年份 2014

【 摘 要 】

Background

For cell therapies to treat diabetes, it is important to produce a sufficient number of pancreatic endocrine cells that function similarly to primary islets. Induced pluripotent stem (iPS) cells represent a potentially unlimited source of functional pancreatic endocrine cells. However, the use of iPS cells for laboratory studies and cell-based therapies is hampered by their high tumorigenic potential and limited ability to generate pure populations of differentiated cell types in vitro. The purpose of this study was to establish a pancreatic stem cell line from iPS cells derived from mouse fibroblasts.

Methods

Mouse iPS cells were induced to differentiate into insulin-producing cells by a multi-step differentiation protocol, which was conducted as described previously with minor modifications. Selection of the pancreatic stem cell was based on morphology and Pdx1 expression. The pancreatic potential of the pancreatic stem cells was evaluated using a reverse transcription PCR, real-time PCR, immunofluorescence, and a glucose challenge test. To assess potential tumorigenicity of the pancreatic stem cells, the cells were injected into the quadriceps femoris muscle of the left hindlimb of nude mice.

Results

The iPS-derived pancreatic stem cells expressed the transcription factor –Pdx1– a marker of pancreatic development, and continued to divide actively beyond passage 80. Endocrine cells derived from these pancreatic stem cells expressed insulin and pancreatic genes, and they released insulin in response to glucose stimulation. Mice injected with the pancreatic stem cells did not develop tumors, in contrast to mice injected with an equal number of iPS cells.

Conclusion

This strategy provides a new approach for generation of insulin-producing cells that is more efficient and safer than using iPS cells. We believe that this approach will help to develop a patient-specific cell transplantation therapy for diabetes in the near future.

【 授权许可】

   
2014 Kuise et al.; licensee BioMed Central Ltd.

【 图 表 】

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