Gingival fibroblasts as a promising source of induced pluripotent stem cells.

Hiroshi Egusa, Keisuke Okita, Hiroki Kayashima, Guannan Yu, Sho Fukuyasu, Makio Saeki, Takuya Matsumoto, Shinya Yamanaka, Hirofumi Yatani

Research output: Contribution to journalArticle

87 Citations (Scopus)

Abstract

Induced pluripotent stem (iPS) cells efficiently generated from accessible tissues have the potential for clinical applications. Oral gingiva, which is often resected during general dental treatments and treated as biomedical waste, is an easily obtainable tissue, and cells can be isolated from patients with minimal discomfort. We herein demonstrate iPS cell generation from adult wild-type mouse gingival fibroblasts (GFs) via introduction of four factors (Oct3/4, Sox2, Klf4 and c-Myc; GF-iPS-4F cells) or three factors (the same as GF-iPS-4F cells, but without the c-Myc oncogene; GF-iPS-3F cells) without drug selection. iPS cells were also generated from primary human gingival fibroblasts via four-factor transduction. These cells exhibited the morphology and growth properties of embryonic stem (ES) cells and expressed ES cell marker genes, with a decreased CpG methylation ratio in promoter regions of Nanog and Oct3/4. Additionally, teratoma formation assays showed ES cell-like derivation of cells and tissues representative of all three germ layers. In comparison to mouse GF-iPS-4F cells, GF-iPS-3F cells showed consistently more ES cell-like characteristics in terms of DNA methylation status and gene expression, although the reprogramming process was substantially delayed and the overall efficiency was also reduced. When transplanted into blastocysts, GF-iPS-3F cells gave rise to chimeras and contributed to the development of the germline. Notably, the four-factor reprogramming efficiency of mouse GFs was more than 7-fold higher than that of fibroblasts from tail-tips, possibly because of their high proliferative capacity. These results suggest that GFs from the easily obtainable gingival tissues can be readily reprogrammed into iPS cells, thus making them a promising cell source for investigating the basis of cellular reprogramming and pluripotency for future clinical applications. In addition, high-quality iPS cells were generated from mouse GFs without Myc transduction or a specific system for reprogrammed cell selection.

Original languageEnglish
Article numbere12743
JournalPLoS One
Volume5
Issue number9
DOIs
Publication statusPublished - 2010
Externally publishedYes

Fingerprint

Induced Pluripotent Stem Cells
Fibroblasts
Stem cells
fibroblasts
embryonic stem cells
Embryonic Stem Cells
Tissue
mice
cells
induced pluripotent stem cells
gingiva
Germ Layers
myc Genes
chimerism
oncogenes
Teratoma
Gingiva
Blastocyst
DNA methylation
DNA Methylation

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Egusa, H., Okita, K., Kayashima, H., Yu, G., Fukuyasu, S., Saeki, M., ... Yatani, H. (2010). Gingival fibroblasts as a promising source of induced pluripotent stem cells. PLoS One, 5(9), [e12743]. https://doi.org/10.1371/journal.pone.0012743

Gingival fibroblasts as a promising source of induced pluripotent stem cells. / Egusa, Hiroshi; Okita, Keisuke; Kayashima, Hiroki; Yu, Guannan; Fukuyasu, Sho; Saeki, Makio; Matsumoto, Takuya; Yamanaka, Shinya; Yatani, Hirofumi.

In: PLoS One, Vol. 5, No. 9, e12743, 2010.

Research output: Contribution to journalArticle

Egusa, H, Okita, K, Kayashima, H, Yu, G, Fukuyasu, S, Saeki, M, Matsumoto, T, Yamanaka, S & Yatani, H 2010, 'Gingival fibroblasts as a promising source of induced pluripotent stem cells.', PLoS One, vol. 5, no. 9, e12743. https://doi.org/10.1371/journal.pone.0012743
Egusa H, Okita K, Kayashima H, Yu G, Fukuyasu S, Saeki M et al. Gingival fibroblasts as a promising source of induced pluripotent stem cells. PLoS One. 2010;5(9). e12743. https://doi.org/10.1371/journal.pone.0012743
Egusa, Hiroshi ; Okita, Keisuke ; Kayashima, Hiroki ; Yu, Guannan ; Fukuyasu, Sho ; Saeki, Makio ; Matsumoto, Takuya ; Yamanaka, Shinya ; Yatani, Hirofumi. / Gingival fibroblasts as a promising source of induced pluripotent stem cells. In: PLoS One. 2010 ; Vol. 5, No. 9.
@article{4ead586e52614db587369c80fb8d923a,
title = "Gingival fibroblasts as a promising source of induced pluripotent stem cells.",
abstract = "Induced pluripotent stem (iPS) cells efficiently generated from accessible tissues have the potential for clinical applications. Oral gingiva, which is often resected during general dental treatments and treated as biomedical waste, is an easily obtainable tissue, and cells can be isolated from patients with minimal discomfort. We herein demonstrate iPS cell generation from adult wild-type mouse gingival fibroblasts (GFs) via introduction of four factors (Oct3/4, Sox2, Klf4 and c-Myc; GF-iPS-4F cells) or three factors (the same as GF-iPS-4F cells, but without the c-Myc oncogene; GF-iPS-3F cells) without drug selection. iPS cells were also generated from primary human gingival fibroblasts via four-factor transduction. These cells exhibited the morphology and growth properties of embryonic stem (ES) cells and expressed ES cell marker genes, with a decreased CpG methylation ratio in promoter regions of Nanog and Oct3/4. Additionally, teratoma formation assays showed ES cell-like derivation of cells and tissues representative of all three germ layers. In comparison to mouse GF-iPS-4F cells, GF-iPS-3F cells showed consistently more ES cell-like characteristics in terms of DNA methylation status and gene expression, although the reprogramming process was substantially delayed and the overall efficiency was also reduced. When transplanted into blastocysts, GF-iPS-3F cells gave rise to chimeras and contributed to the development of the germline. Notably, the four-factor reprogramming efficiency of mouse GFs was more than 7-fold higher than that of fibroblasts from tail-tips, possibly because of their high proliferative capacity. These results suggest that GFs from the easily obtainable gingival tissues can be readily reprogrammed into iPS cells, thus making them a promising cell source for investigating the basis of cellular reprogramming and pluripotency for future clinical applications. In addition, high-quality iPS cells were generated from mouse GFs without Myc transduction or a specific system for reprogrammed cell selection.",
author = "Hiroshi Egusa and Keisuke Okita and Hiroki Kayashima and Guannan Yu and Sho Fukuyasu and Makio Saeki and Takuya Matsumoto and Shinya Yamanaka and Hirofumi Yatani",
year = "2010",
doi = "10.1371/journal.pone.0012743",
language = "English",
volume = "5",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "9",

}

TY - JOUR

T1 - Gingival fibroblasts as a promising source of induced pluripotent stem cells.

AU - Egusa, Hiroshi

AU - Okita, Keisuke

AU - Kayashima, Hiroki

AU - Yu, Guannan

AU - Fukuyasu, Sho

AU - Saeki, Makio

AU - Matsumoto, Takuya

AU - Yamanaka, Shinya

AU - Yatani, Hirofumi

PY - 2010

Y1 - 2010

N2 - Induced pluripotent stem (iPS) cells efficiently generated from accessible tissues have the potential for clinical applications. Oral gingiva, which is often resected during general dental treatments and treated as biomedical waste, is an easily obtainable tissue, and cells can be isolated from patients with minimal discomfort. We herein demonstrate iPS cell generation from adult wild-type mouse gingival fibroblasts (GFs) via introduction of four factors (Oct3/4, Sox2, Klf4 and c-Myc; GF-iPS-4F cells) or three factors (the same as GF-iPS-4F cells, but without the c-Myc oncogene; GF-iPS-3F cells) without drug selection. iPS cells were also generated from primary human gingival fibroblasts via four-factor transduction. These cells exhibited the morphology and growth properties of embryonic stem (ES) cells and expressed ES cell marker genes, with a decreased CpG methylation ratio in promoter regions of Nanog and Oct3/4. Additionally, teratoma formation assays showed ES cell-like derivation of cells and tissues representative of all three germ layers. In comparison to mouse GF-iPS-4F cells, GF-iPS-3F cells showed consistently more ES cell-like characteristics in terms of DNA methylation status and gene expression, although the reprogramming process was substantially delayed and the overall efficiency was also reduced. When transplanted into blastocysts, GF-iPS-3F cells gave rise to chimeras and contributed to the development of the germline. Notably, the four-factor reprogramming efficiency of mouse GFs was more than 7-fold higher than that of fibroblasts from tail-tips, possibly because of their high proliferative capacity. These results suggest that GFs from the easily obtainable gingival tissues can be readily reprogrammed into iPS cells, thus making them a promising cell source for investigating the basis of cellular reprogramming and pluripotency for future clinical applications. In addition, high-quality iPS cells were generated from mouse GFs without Myc transduction or a specific system for reprogrammed cell selection.

AB - Induced pluripotent stem (iPS) cells efficiently generated from accessible tissues have the potential for clinical applications. Oral gingiva, which is often resected during general dental treatments and treated as biomedical waste, is an easily obtainable tissue, and cells can be isolated from patients with minimal discomfort. We herein demonstrate iPS cell generation from adult wild-type mouse gingival fibroblasts (GFs) via introduction of four factors (Oct3/4, Sox2, Klf4 and c-Myc; GF-iPS-4F cells) or three factors (the same as GF-iPS-4F cells, but without the c-Myc oncogene; GF-iPS-3F cells) without drug selection. iPS cells were also generated from primary human gingival fibroblasts via four-factor transduction. These cells exhibited the morphology and growth properties of embryonic stem (ES) cells and expressed ES cell marker genes, with a decreased CpG methylation ratio in promoter regions of Nanog and Oct3/4. Additionally, teratoma formation assays showed ES cell-like derivation of cells and tissues representative of all three germ layers. In comparison to mouse GF-iPS-4F cells, GF-iPS-3F cells showed consistently more ES cell-like characteristics in terms of DNA methylation status and gene expression, although the reprogramming process was substantially delayed and the overall efficiency was also reduced. When transplanted into blastocysts, GF-iPS-3F cells gave rise to chimeras and contributed to the development of the germline. Notably, the four-factor reprogramming efficiency of mouse GFs was more than 7-fold higher than that of fibroblasts from tail-tips, possibly because of their high proliferative capacity. These results suggest that GFs from the easily obtainable gingival tissues can be readily reprogrammed into iPS cells, thus making them a promising cell source for investigating the basis of cellular reprogramming and pluripotency for future clinical applications. In addition, high-quality iPS cells were generated from mouse GFs without Myc transduction or a specific system for reprogrammed cell selection.

UR - http://www.scopus.com/inward/record.url?scp=77958592067&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77958592067&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0012743

DO - 10.1371/journal.pone.0012743

M3 - Article

C2 - 20856871

AN - SCOPUS:77958592067

VL - 5

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 9

M1 - e12743

ER -