Human Cell

, Volume 20, Issue 3, pp 77–84

Differentiation of mesenchymal cells derived from human amniotic membranes into hepatocyte-like cells in vitro

  • Tomoharu Tamagawa
  • Satoshi Oi
  • Isamu Ishiwata
  • Hiroshi Ishikawa
  • Yukio Nakamura
Original Article


Mesenchymal stem cells are believed to be involved in the formation of mesenchymal tissues, including bone, cartilage, muscle, tendon and adipose tissue. Interestingly, it has previously been reported that mesenchymal stem cells could also differentiate into endoderm-derived cells, such as hepatocytes. The amniotic membrane contains mesenchymal cells and is a readily available human tissue. Therefore, we investigated the potential of mesenchymal cells derived from human amniotic membrane (MC-HAM) to differentiate into hepatocytes. We analyzed the expression of hepatocyte-specific genes in MC-HAM before and after induction of differentiation into hepatocytes. We observed the expression of mRNAs encoding albumin, a-fetoprotein, cytokeratin 18 and α1-antitrypsin, but not those encoding glucose-6-phosphatase or ornithine transcarbamylase, prior to the induction of differentiation. However, immunocytochemistry revealed that albumin and α-fetoprotein were abundantly produced only afterthe induction of differentiation into hepatocytes. In addition, we observed the storage of glycogen, a characteristic feature of hepatocytes, using periodic acid-Schiff staining of MC-HAM induced to differentiate into hepatocytes. Overall, MC-HAM appear to be able to differentiate into cells possessing some characteristics of hepatocytes. Although further studies should be carried out to determine whether such in vitro-differentiated cells can function in vivo as hepatocytes. These cells may be useful in various applications that require human hepatocytes.

Key words

amnion membrane differentiation hepatocyte-like cell mesanchymal stemcell regeneration medicine 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Kamlot A, Rozga J, Watanabe FD et al. Artificial liver support systems. Biotecnol Bioeng 1995; 50: 382–91.CrossRefGoogle Scholar
  2. 2.
    Rambhatla L, Chiu CP, Kundu P et al. Generation of hepatocyte-like cells from human embryonic stem cells. Cell Transplant 2003; 12: 1–11.PubMedCrossRefGoogle Scholar
  3. 3.
    Danet GH, Luongo JL, Butler G et al. C1qRp defines a new human stem cell population with hematopoietic and hepatic potential. Proc Natl Acad Sci USA 2002; 99: 10441–5.PubMedCrossRefGoogle Scholar
  4. 4.
    Beerheide W, von Mach MA, Ringel M et al. Downregulation of β2-microglobulin in human cord blood somatic stem cells after transplantation into livers of SCID-mice: an escape mechanism of stem cells? Biochem Biophys Res Commun 2002; 294: 1052–63.PubMedCrossRefGoogle Scholar
  5. 5.
    Kakinuma S, Tanaka Y, Chinzei R et al. Human umbilical cord blood as a source of transplantable hepatic progenitor cells. Stem Cells 2003; 21: 217–27.PubMedCrossRefGoogle Scholar
  6. 6.
    Takashima S, Ise H, Zhao P et al. Human amnion epithelial cells possess hepatocyte-like characteristics and function. Cell Struc Func 2004; 29: 73–84.CrossRefGoogle Scholar
  7. 7.
    Whittle WL, Gibb W, Challis JRG. The characterization of human amnion epithelial and mesenchymal cells: the cellular expression, activity and glucocorticoid regulation of prostaglandin output. Placenta 2000; 21: 394–401.PubMedCrossRefGoogle Scholar
  8. 8.
    Sakuragawa N, Kakinuma K, Kikuchi A et al. Human amnion mesenchyme cells express phenotypes of neuroglial progenitor cells. J Neurosci Res 2004; 78: 208–14.PubMedCrossRefGoogle Scholar
  9. 9.
    Pittenger MF, Mackay AM, Beck SC et al. Multilineage potential of adult human mesenchymal stem cells. Science 1999; 284: 143–7.PubMedCrossRefGoogle Scholar
  10. 10.
    Tamagawa T, Ishiwata I, Saito S. Establishment and characterization of a pluripotent stem cell line derived from human amniotic membranes and initiation of germ layers in vitro. Human Cell 2004; 17: 125–30.PubMedCrossRefGoogle Scholar
  11. 11.
    Tamagawa T, Ishiwata I, Nakamura Y. Differentiation of human amniotic membrane cells into osteoblasts in vitro. Human Cell 2006; 18: 191–5.CrossRefGoogle Scholar
  12. 12.
    Ishiwata I, Ishiwata C, Soma M et al. Establishment and characterization of HUOT, a human ovarian malignant teratoma cell line producing alpha-fetoprotein. J Natl Cancer Inst 1985; 75: 411–22.PubMedGoogle Scholar
  13. 13.
    Michalopoulos GK, DeFrances MC. Liver regeneration. Science 1997; 276: 60–6.PubMedCrossRefGoogle Scholar
  14. 14.
    Oh SH, Miyazaki M, Kouchi H et al. Hepatocyte growth factor induces differentiation of adult rat bone marrow cells into a hepatocyte lineage in vitro. Biochem Biophys Res Commun 2000; 279: 500–04.PubMedCrossRefGoogle Scholar
  15. 15.
    Jung J, Zheng M, Goldfarb M et al. Initiation of mammalian liver development from endoderm by fibroblast growth factor. Science 1999; 284: 1998–2003.PubMedCrossRefGoogle Scholar
  16. 16.
    Kamiya A, Kinoshita T, Ito Y et al. Fetal liver development requires a paracrine action of oncostatin M through the gp130 signal transducer. EMBO J 1999; 18: 2127–36.PubMedCrossRefGoogle Scholar
  17. 17.
    Koide N, Shinji T, Tanabe T et al. Continued high albumin production by multicellular spheroids of adult rat hepatocytes formed in the presence of liver-derived proteoglycans. Biochem Biophys Res Commun 1989; 161: 385–91.PubMedCrossRefGoogle Scholar
  18. 18.
    Mooney DJ, Langer R, Ingber DE. Cytoskeletal filament assembly and the control of cell spreading and function by extracellular matrix. J Cell Sci 1995; 108: 2311–20.PubMedGoogle Scholar
  19. 19.
    Sanchez A. Fibronectin regulates morphology, cell organization and gene expression of rat fetal hepatocytes in primary culture. J Hepatol 2000; 32: 242–50.PubMedCrossRefGoogle Scholar
  20. 20.
    Chien CC, Yen BL, Lee FK et al. In vitro differentiation of human placenta-derived multipotent cells into hepatocytelike cells. Stem Cells 2006; 24: 1759–68.PubMedCrossRefGoogle Scholar
  21. 21.
    Parviz Z, Matullo C, Garrison WD et al. Hepatocyte nuclear factor 4α controls the development of a hepatic epithelium and liver morphogenesis. Nat Genet 2003; 34: 292–6.PubMedCrossRefGoogle Scholar
  22. 22.
    Sakuragawa N, Enosawa S, Ishii T et al. Human amniotic epithelial cells are promising transgene carriers for allogeneic cell transplantation into liver. J Hum Gene 2000; 45: 171–6.CrossRefGoogle Scholar

Copyright information

© Society and Springer Japan 2007

Authors and Affiliations

  • Tomoharu Tamagawa
    • 1
    • 2
  • Satoshi Oi
    • 3
  • Isamu Ishiwata
    • 1
  • Hiroshi Ishikawa
    • 3
  • Yukio Nakamura
    • 2
  1. 1.Institute of Cell BiologyIshiwata HospitalJapan
  2. 2.Cell Engineering DivisionRIKEN BioResource CenterIbarakiJapan
  3. 3.Department of AnatomyJikei University School of MedicineTokyoJapan

Personalised recommendations