Generation of functional insulin-producing cells from mouse embryonic stem cells through 804G cell-derived extracellular matrix and protein transduction of transcription factors

Taku Kaitsuka, Hirofumi Noguchi, Nobuaki Shiraki, Takuya Kubo, Fan Yan Wei, Farzana Hakim, Shoen Kume, Kazuhito Tomizawa

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Embryonic stem (ES) and induced pluripotent stem (iPS) cells have potential applications to regenerative medicine for diabetes; however, a useful and safe way to generate pancreatic β cells has not been developed. In this study, we tried to establish an effective method of differentiation through the protein transduction of three transcription factors (Pdx1, NeuroD, and MafA) important to pancreatic β cell development. The method poses no risk of unexpected genetic modifications in target cells. Transduction of the three proteins induced the differentiation of mouse ES and mouse iPS cells into insulin-producing cells. Furthermore, a laminin-5-rich extracellular matrix efficiently induced differentiation under feeder-free conditions. Cell differentiation was confirmed with the expression of the insulin 1 gene in addition to marker genes in pancreatic β cells, the differentiated cells secreted glucose-responsive C-peptide, and their transplantation restored normoglycemia in diabetic mice. Moreover, Pdx1 protein transduction had facilitative effects on differentiation into pancreatic endocrine progenitors from human iPS cells. These results suggest the direct delivery of recombinant proteins and treatment with laminin-5-rich extracellular matrix to be useful for the generation of insulinproducing cells.

Original languageEnglish
Pages (from-to)114-127
Number of pages14
JournalStem cells translational medicine
Volume3
Issue number1
DOIs
Publication statusPublished - Jan 1 2014
Externally publishedYes

Fingerprint

Extracellular Matrix Proteins
Transcription Factors
Insulin
Induced Pluripotent Stem Cells
Extracellular Matrix
Proteins
Regenerative Medicine
C-Peptide
Mouse Embryonic Stem Cells
Recombinant Proteins
Genes
Cell Differentiation
Transplantation
Glucose

Keywords

  • Diabetes
  • Embryonic stem cells
  • Induced pluripotent stem cells
  • Pancreatic differentiation
  • Transcription factors

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

Cite this

Generation of functional insulin-producing cells from mouse embryonic stem cells through 804G cell-derived extracellular matrix and protein transduction of transcription factors. / Kaitsuka, Taku; Noguchi, Hirofumi; Shiraki, Nobuaki; Kubo, Takuya; Wei, Fan Yan; Hakim, Farzana; Kume, Shoen; Tomizawa, Kazuhito.

In: Stem cells translational medicine, Vol. 3, No. 1, 01.01.2014, p. 114-127.

Research output: Contribution to journalArticle

Kaitsuka, Taku ; Noguchi, Hirofumi ; Shiraki, Nobuaki ; Kubo, Takuya ; Wei, Fan Yan ; Hakim, Farzana ; Kume, Shoen ; Tomizawa, Kazuhito. / Generation of functional insulin-producing cells from mouse embryonic stem cells through 804G cell-derived extracellular matrix and protein transduction of transcription factors. In: Stem cells translational medicine. 2014 ; Vol. 3, No. 1. pp. 114-127.
@article{21cf934231174cbc99cb49467f47ba9d,
title = "Generation of functional insulin-producing cells from mouse embryonic stem cells through 804G cell-derived extracellular matrix and protein transduction of transcription factors",
abstract = "Embryonic stem (ES) and induced pluripotent stem (iPS) cells have potential applications to regenerative medicine for diabetes; however, a useful and safe way to generate pancreatic β cells has not been developed. In this study, we tried to establish an effective method of differentiation through the protein transduction of three transcription factors (Pdx1, NeuroD, and MafA) important to pancreatic β cell development. The method poses no risk of unexpected genetic modifications in target cells. Transduction of the three proteins induced the differentiation of mouse ES and mouse iPS cells into insulin-producing cells. Furthermore, a laminin-5-rich extracellular matrix efficiently induced differentiation under feeder-free conditions. Cell differentiation was confirmed with the expression of the insulin 1 gene in addition to marker genes in pancreatic β cells, the differentiated cells secreted glucose-responsive C-peptide, and their transplantation restored normoglycemia in diabetic mice. Moreover, Pdx1 protein transduction had facilitative effects on differentiation into pancreatic endocrine progenitors from human iPS cells. These results suggest the direct delivery of recombinant proteins and treatment with laminin-5-rich extracellular matrix to be useful for the generation of insulinproducing cells.",
keywords = "Diabetes, Embryonic stem cells, Induced pluripotent stem cells, Pancreatic differentiation, Transcription factors",
author = "Taku Kaitsuka and Hirofumi Noguchi and Nobuaki Shiraki and Takuya Kubo and Wei, {Fan Yan} and Farzana Hakim and Shoen Kume and Kazuhito Tomizawa",
year = "2014",
month = "1",
day = "1",
doi = "10.5966/sctm.2013-0075",
language = "English",
volume = "3",
pages = "114--127",
journal = "Stem cells translational medicine",
issn = "2157-6564",
publisher = "AlphaMed Press",
number = "1",

}

TY - JOUR

T1 - Generation of functional insulin-producing cells from mouse embryonic stem cells through 804G cell-derived extracellular matrix and protein transduction of transcription factors

AU - Kaitsuka, Taku

AU - Noguchi, Hirofumi

AU - Shiraki, Nobuaki

AU - Kubo, Takuya

AU - Wei, Fan Yan

AU - Hakim, Farzana

AU - Kume, Shoen

AU - Tomizawa, Kazuhito

PY - 2014/1/1

Y1 - 2014/1/1

N2 - Embryonic stem (ES) and induced pluripotent stem (iPS) cells have potential applications to regenerative medicine for diabetes; however, a useful and safe way to generate pancreatic β cells has not been developed. In this study, we tried to establish an effective method of differentiation through the protein transduction of three transcription factors (Pdx1, NeuroD, and MafA) important to pancreatic β cell development. The method poses no risk of unexpected genetic modifications in target cells. Transduction of the three proteins induced the differentiation of mouse ES and mouse iPS cells into insulin-producing cells. Furthermore, a laminin-5-rich extracellular matrix efficiently induced differentiation under feeder-free conditions. Cell differentiation was confirmed with the expression of the insulin 1 gene in addition to marker genes in pancreatic β cells, the differentiated cells secreted glucose-responsive C-peptide, and their transplantation restored normoglycemia in diabetic mice. Moreover, Pdx1 protein transduction had facilitative effects on differentiation into pancreatic endocrine progenitors from human iPS cells. These results suggest the direct delivery of recombinant proteins and treatment with laminin-5-rich extracellular matrix to be useful for the generation of insulinproducing cells.

AB - Embryonic stem (ES) and induced pluripotent stem (iPS) cells have potential applications to regenerative medicine for diabetes; however, a useful and safe way to generate pancreatic β cells has not been developed. In this study, we tried to establish an effective method of differentiation through the protein transduction of three transcription factors (Pdx1, NeuroD, and MafA) important to pancreatic β cell development. The method poses no risk of unexpected genetic modifications in target cells. Transduction of the three proteins induced the differentiation of mouse ES and mouse iPS cells into insulin-producing cells. Furthermore, a laminin-5-rich extracellular matrix efficiently induced differentiation under feeder-free conditions. Cell differentiation was confirmed with the expression of the insulin 1 gene in addition to marker genes in pancreatic β cells, the differentiated cells secreted glucose-responsive C-peptide, and their transplantation restored normoglycemia in diabetic mice. Moreover, Pdx1 protein transduction had facilitative effects on differentiation into pancreatic endocrine progenitors from human iPS cells. These results suggest the direct delivery of recombinant proteins and treatment with laminin-5-rich extracellular matrix to be useful for the generation of insulinproducing cells.

KW - Diabetes

KW - Embryonic stem cells

KW - Induced pluripotent stem cells

KW - Pancreatic differentiation

KW - Transcription factors

UR - http://www.scopus.com/inward/record.url?scp=84892689842&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84892689842&partnerID=8YFLogxK

U2 - 10.5966/sctm.2013-0075

DO - 10.5966/sctm.2013-0075

M3 - Article

C2 - 24292793

AN - SCOPUS:84892689842

VL - 3

SP - 114

EP - 127

JO - Stem cells translational medicine

JF - Stem cells translational medicine

SN - 2157-6564

IS - 1

ER -