Propagation of adult rat bone marrow-derived hepatocyte-like cells by serial passages in vitro

Masahiro Miyazaki, Takuro Masaka, Ichiro Akiyama, Emiko Nakashima, Masakiyo Sakaguchi, Nam Ho Huh

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Previously, we found that hepatocyte growth factor receptor (c-Met)-and alpha-fetoprotein (AFP)-expressing cells were present in adult rat bone marrow, and that these cells also expressed hematopoietic stem cell markers, such as CD34, Thy-1, and c-Kit. When bone marrow cells were cultured in a hepatocyte growth medium (HGM) with HGF and EGF, colonies composed of polygonal cells resembling mature hepatocytes appeared by 2 weeks and grew very slowly because of overgrowth of stromal cells. At days 34-41, 2-mm2 sheets of hepatocyte-like cells were cut out of their colonies by scratching with an injection needle under observation with a phase contrast microscope, transferred into wells of 24-well plates, and cultured in the HGM medium in the presence or absence of HGF and EGF. When cells reached confluence, cells were detached with trypsin and EDTA and transferred step by step into bigger culture vessels. Thus, hepatocyte-like cells were expanded 1000-fold during less than 4 months. These cells were immunocytochemically stained for albumin and also for AFP and the hematopoietic stem cell markers described above, showing characteristics of oval cells. By RT-PCR, we detected mRNAs of tryptophan-2,3-dioxygenase and tyrosine aminotransferase, markers of hepatocytes at a terminal differentiation stage. The present culture system may be useful for supply of hepatocyte resources for cell transplantation therapy.

Original languageEnglish
Pages (from-to)385-391
Number of pages7
JournalCell Transplantation
Volume13
Issue number4
Publication statusPublished - 2004

Fingerprint

Serial Passage
Rats
Hepatocytes
Bone
Bone Marrow
alpha-Fetoproteins
Hematopoietic Stem Cells
Stem cells
Epidermal Growth Factor
Bone Marrow Cells
Proto-Oncogene Proteins c-met
Tyrosine Transaminase
In Vitro Techniques
Cell Transplantation
Ethylenediaminetetraacetic acid
Stromal Cells
Growth
Cell- and Tissue-Based Therapy
Edetic Acid
Tryptophan

Keywords

  • Albumin
  • Bone marrow cells
  • Hepatocyte-like cells
  • In vitro expansion
  • Tryptophan-2,3-dioxygenase
  • Tyrosine aminotransferase

ASJC Scopus subject areas

  • Cell Biology
  • Transplantation

Cite this

Miyazaki, M., Masaka, T., Akiyama, I., Nakashima, E., Sakaguchi, M., & Huh, N. H. (2004). Propagation of adult rat bone marrow-derived hepatocyte-like cells by serial passages in vitro. Cell Transplantation, 13(4), 385-391.

Propagation of adult rat bone marrow-derived hepatocyte-like cells by serial passages in vitro. / Miyazaki, Masahiro; Masaka, Takuro; Akiyama, Ichiro; Nakashima, Emiko; Sakaguchi, Masakiyo; Huh, Nam Ho.

In: Cell Transplantation, Vol. 13, No. 4, 2004, p. 385-391.

Research output: Contribution to journalArticle

Miyazaki, M, Masaka, T, Akiyama, I, Nakashima, E, Sakaguchi, M & Huh, NH 2004, 'Propagation of adult rat bone marrow-derived hepatocyte-like cells by serial passages in vitro', Cell Transplantation, vol. 13, no. 4, pp. 385-391.
Miyazaki, Masahiro ; Masaka, Takuro ; Akiyama, Ichiro ; Nakashima, Emiko ; Sakaguchi, Masakiyo ; Huh, Nam Ho. / Propagation of adult rat bone marrow-derived hepatocyte-like cells by serial passages in vitro. In: Cell Transplantation. 2004 ; Vol. 13, No. 4. pp. 385-391.
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