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

Takashi Kuise, Hirofumi Noguchi, Hiroshi Tazawa, Takashi Kawai, Masaya Iwamuro, Issei Saitoh, Hitomi Kataoka, Masami Watanabe, Yasufumi Noguchi, Toshiyoshi Fujiwara

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

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.

Original languageEnglish
Article number64
JournalBioMedical Engineering Online
Volume13
Issue number1
DOIs
Publication statusPublished - May 27 2014

Fingerprint

Induced Pluripotent Stem Cells
Fibroblasts
Stem cells
Stem Cells
Cell Line
Endocrine Cells
Cell- and Tissue-Based Therapy
Insulin
Quadriceps Muscle
Glucose
Cells
Medical problems
Cell Transplantation
Hindlimb
Nude Mice
Reverse Transcription
Fluorescent Antibody Technique
Real-Time Polymerase Chain Reaction
Transcription Factors
Transcription factors

Keywords

  • Diabetes
  • iPS cells
  • Mouse pancreatic stem cells

ASJC Scopus subject areas

  • Biomedical Engineering
  • Radiological and Ultrasound Technology
  • Radiology Nuclear Medicine and imaging
  • Biomaterials

Cite this

Establishment of a pancreatic stem cell line from fibroblast-derived induced pluripotent stem cells. / Kuise, Takashi; Noguchi, Hirofumi; Tazawa, Hiroshi; Kawai, Takashi; Iwamuro, Masaya; Saitoh, Issei; Kataoka, Hitomi; Watanabe, Masami; Noguchi, Yasufumi; Fujiwara, Toshiyoshi.

In: BioMedical Engineering Online, Vol. 13, No. 1, 64, 27.05.2014.

Research output: Contribution to journalArticle

@article{30292037237342889d59a6b89e0946d7,
title = "Establishment of a pancreatic stem cell line from fibroblast-derived induced pluripotent stem cells",
abstract = "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.",
keywords = "Diabetes, iPS cells, Mouse pancreatic stem cells",
author = "Takashi Kuise and Hirofumi Noguchi and Hiroshi Tazawa and Takashi Kawai and Masaya Iwamuro and Issei Saitoh and Hitomi Kataoka and Masami Watanabe and Yasufumi Noguchi and Toshiyoshi Fujiwara",
year = "2014",
month = "5",
day = "27",
doi = "10.1186/1475-925X-13-64",
language = "English",
volume = "13",
journal = "BioMedical Engineering Online",
issn = "1475-925X",
publisher = "BioMed Central",
number = "1",

}

TY - JOUR

T1 - Establishment of a pancreatic stem cell line from fibroblast-derived induced pluripotent stem cells

AU - Kuise, Takashi

AU - Noguchi, Hirofumi

AU - Tazawa, Hiroshi

AU - Kawai, Takashi

AU - Iwamuro, Masaya

AU - Saitoh, Issei

AU - Kataoka, Hitomi

AU - Watanabe, Masami

AU - Noguchi, Yasufumi

AU - Fujiwara, Toshiyoshi

PY - 2014/5/27

Y1 - 2014/5/27

N2 - 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.

AB - 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.

KW - Diabetes

KW - iPS cells

KW - Mouse pancreatic stem cells

UR - http://www.scopus.com/inward/record.url?scp=84902285427&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84902285427&partnerID=8YFLogxK

U2 - 10.1186/1475-925X-13-64

DO - 10.1186/1475-925X-13-64

M3 - Article

C2 - 24886514

AN - SCOPUS:84902285427

VL - 13

JO - BioMedical Engineering Online

JF - BioMedical Engineering Online

SN - 1475-925X

IS - 1

M1 - 64

ER -