Comparative analysis of endoderm formation efficiency between mouse ES cells and iPS cells

Masaya Iwamuro, Toshiyuki Komaki, Yasuhiro Kubota, Masayuki Seita, Hironobu Kawamoto, Takeshi Yuasa, Javed M. Shahid, Reham A R A Hassan, Wael A R A Hassan, Shuhei Nakaji, Yuriko Nishikawa, Eisaku Kondo, Kazuhide Yamamoto, Naoya Kobayashi

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Definitive endoderm (DE) derived from stem cells holds potential to differentiate into hepatocytes. Stem cell therapy using those cells has potential for a treatment of liver disease. To date, various ways of inducing hepatocytes from embryonic stem (ES) cells have been reported by researchers. However, it has not been proved enough that induced pluripotent stem (iPS) cells behave in the same manner as ES cells in endoderm differentiation. The purpose of this study was to establish an efficient method to induce DE from iPS cells, through comparatively analyzing the efficacy of endoderm formation from mouse ES cells. Furthermore, the efficiency of a serum-free medium in the differentiation into DE was investigated. Mouse ES cells and iPS cells were floated in culture medium for 2 or 5 days and embryoid bodies (EB) were formed. Subsequently, DE was induced with 100 ng/ml activin A and 100 ng/ml basic fibroblast growth factor (bFGF). RT-PCR and real-time PCR analyses were carried out at each step to determine the gene expression of EB markers. The difference in cellular proliferation between serum-containing and serum-free media was examined by an MTS assay in EB and DE induction. iPS cells showed the paralleled mRNA expression to ES cells in each step of differentiation into EB, but the levels of expression of Sox17 and Foxa2 were relatively higher in ES cell-derived DE, whereas Cxcr4 expression was higher in iPS cell-derived DE. The utilization of serum-free medium for iPS cells showed significantly favorable cellular proliferation during EB formation and subsequent DE induction. Forming EB for 5 days and subsequently DE induction with activin A and bFGF with serum-free medium was an appropriate protocol in iPS cells. This may represent an important step for generating hepatocytes from iPS cells for the development of cell therapy.

Original languageEnglish
Pages (from-to)831-839
Number of pages9
JournalCell Transplantation
Volume19
Issue number6-7
DOIs
Publication statusPublished - 2010

Fingerprint

Induced Pluripotent Stem Cells
Endoderm
Stem cells
Embryoid Bodies
Serum-Free Culture Media
Embryonic Stem Cells
Hepatocytes
Fibroblast Growth Factor 2
Cell- and Tissue-Based Therapy
Mouse Embryonic Stem Cells
Stem Cells
Cell Proliferation
Fibroblasts
Culture Media
Liver Diseases
Real-Time Polymerase Chain Reaction
Research Personnel

Keywords

  • Definitive endoderm (DE)
  • Embryonic stem cells
  • Induced pluripotent stem cells (iPS)

ASJC Scopus subject areas

  • Cell Biology
  • Transplantation
  • Biomedical Engineering

Cite this

Iwamuro, M., Komaki, T., Kubota, Y., Seita, M., Kawamoto, H., Yuasa, T., ... Kobayashi, N. (2010). Comparative analysis of endoderm formation efficiency between mouse ES cells and iPS cells. Cell Transplantation, 19(6-7), 831-839. https://doi.org/10.3727/096368910X508951

Comparative analysis of endoderm formation efficiency between mouse ES cells and iPS cells. / Iwamuro, Masaya; Komaki, Toshiyuki; Kubota, Yasuhiro; Seita, Masayuki; Kawamoto, Hironobu; Yuasa, Takeshi; Shahid, Javed M.; Hassan, Reham A R A; Hassan, Wael A R A; Nakaji, Shuhei; Nishikawa, Yuriko; Kondo, Eisaku; Yamamoto, Kazuhide; Kobayashi, Naoya.

In: Cell Transplantation, Vol. 19, No. 6-7, 2010, p. 831-839.

Research output: Contribution to journalArticle

Iwamuro, M, Komaki, T, Kubota, Y, Seita, M, Kawamoto, H, Yuasa, T, Shahid, JM, Hassan, RARA, Hassan, WARA, Nakaji, S, Nishikawa, Y, Kondo, E, Yamamoto, K & Kobayashi, N 2010, 'Comparative analysis of endoderm formation efficiency between mouse ES cells and iPS cells', Cell Transplantation, vol. 19, no. 6-7, pp. 831-839. https://doi.org/10.3727/096368910X508951
Iwamuro, Masaya ; Komaki, Toshiyuki ; Kubota, Yasuhiro ; Seita, Masayuki ; Kawamoto, Hironobu ; Yuasa, Takeshi ; Shahid, Javed M. ; Hassan, Reham A R A ; Hassan, Wael A R A ; Nakaji, Shuhei ; Nishikawa, Yuriko ; Kondo, Eisaku ; Yamamoto, Kazuhide ; Kobayashi, Naoya. / Comparative analysis of endoderm formation efficiency between mouse ES cells and iPS cells. In: Cell Transplantation. 2010 ; Vol. 19, No. 6-7. pp. 831-839.
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