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Cell and Tissue Research

, Volume 339, Issue 3, pp 505–512 | Cite as

In vitro hepatic maturation of human embryonic stem cells by using a mesenchymal cell line derived from murine fetal livers

  • Takamichi IshiiEmail author
  • Kentaro Yasuchika
  • Ken Fukumitsu
  • Tatsuya Kawamoto
  • Miho Kawamura-Saitoh
  • Yuji Amagai
  • Iwao Ikai
  • Shinji Uemoto
  • Eihachiro Kawase
  • Hirofumi Suemori
  • Norio Nakatsuji
Regular Article

Abstract

Hepatocytes derived from human embryonic stem cells (hESCs) are an attractive cell source for regenerative medicine. We previously reported the differentiation of hESCs into alpha-fetoprotein (AFP)-producing endodermal cells by using extracellular matrix and growth factors. We also reported the establishment of the MLSgt20 cell line, which was derived from mesenchymal cells residing in murine fetal livers and accelerated the hepatic maturation of both murine hepatic progenitor cells and murine ESCs. In this study, hESC-derived AFP-producing cells were isolated by using a flow cytometer and co-cultured with MLSgt20 cells. The co-cultured hESC-derived AFP-producing cells had the immunocytological characteristics of hepatocytes, expressed mature hepatocyte markers (as indicated by reverse transcription and the polymerase chain reaction), and displayed higher hepatocyte functions including ammonia removal, cytochrome P450 3A4/7 activity, and the ability to produce and store glycogen. However, the MLSgt20 cells did not directly cause undifferentiated hESCs to mature into hepatocyte-like cells. The co-culture method was thus successfully shown to induce the differentiation of hESC-derived endodermal cells into functional hepatocyte-like cells.

Keywords

Embryonic stem cells Alpha-fetoprotein Hepatocyte MLSgt20 cell line CYP3A Mouse 

Notes

Acknowledgements

We thank Dr. Andrew G. Farr for providing the anti-gp38 antibodies.

Supplementary material

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Supplementary Fig. 1

(JPEG 119 kb)

441_2009_906_Fig1_ESM.tif (515 kb)
High resolution image (TIFF 515 kb)

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Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • Takamichi Ishii
    • 1
    • 2
    • 7
    Email author
  • Kentaro Yasuchika
    • 2
  • Ken Fukumitsu
    • 1
    • 2
  • Tatsuya Kawamoto
    • 3
  • Miho Kawamura-Saitoh
    • 3
  • Yuji Amagai
    • 3
  • Iwao Ikai
    • 2
    • 4
  • Shinji Uemoto
    • 2
  • Eihachiro Kawase
    • 5
  • Hirofumi Suemori
    • 1
  • Norio Nakatsuji
    • 5
    • 6
  1. 1.Laboratory of Embryonic Stem Cell Research, Stem Cell Research Center, Institute for Frontier Medical SciencesKyoto UniversityKyotoJapan
  2. 2.Department of SurgeryGraduate School of Medicine Kyoto UniversityKyotoJapan
  3. 3.Stem Cell and Drug Discovery InstituteKyotoJapan
  4. 4.Department of Surgery, Kyoto Medical CenterNational Hospital OrganizationKyotoJapan
  5. 5.Department of Development and Differentiation, Institute for Frontier Medical SciencesKyoto UniversityKyotoJapan
  6. 6.Institute for Integrated Cell-Material ScienceKyoto UniversityKyotoJapan
  7. 7.KyotoJapan

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