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Improvement of liver fibrosis by infusion of cultured cells derived from human bone marrow

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Abstract

We develop “autologous bone marrow cell infusion (ABMi) therapy” for the treatment of human decompensated liver cirrhosis and confirm the efficacy and safety of this treatment in multicenter clinical studies. With the goal of further expanding the applications of ABMi, we first cultured human bone marrow cells and then determined whether a cell fraction found to be effective in improving liver fibrosis can be amplified. Cells harvested after two passages (P2 cells) consistently contained approximately 94 % mesenchymal stem cells (MSCs); conversely, the cells harvested after only medium change (P0 cells) contained many macrophages. MSCs (2.8 × 108) in P2 cells were harvested from 3.8 × 108 bone marrow-derived mononuclear cells after 22 days. DNA-chip analysis also showed during the culturing step that bone marrow-derived cells decreased with macrophage phenotype. The infused 5 × 105 P2 cells significantly improved liver fibrosis in the nonobese diabetic/severe combined immunodeficient (NOD-SCID) mouse carbon tetrachloride (CCl4) liver cirrhosis model and induced the expression of matrix metalloproteinase (MMP)-9 and suppressed expressions of alpha smooth muscle actin (αSMA), tumor necrosis factor alpha (TNFα) and transforming growth factor beta (TGFβ) in the liver. Cultured human bone marrow-derived cells (P2 cells) significantly inhibited liver fibrosis. The increase of MMP-9 and suppressed activation of hepatic stellate cells (HSCs) through the regulation of humoral factors (TNFα and TGFβ) contribute to the improvement of liver fibrosis by MSCs comprising about 94 % of P2 cells. MSCs in cultured human bone marrow-derived mono-nuclear cells (BM-MNCs) proliferate sufficiently in cell therapy, so we believe our cultured bone marrow-derived cell therapy can lead to expanded clinical applications and enable outpatient therapy.

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Abbreviations

αSMA:

Alpha smooth muscle actin

ABMi:

Autologous bone marrow cell infusion

BM-MNC:

Bone marrow-derived mononuclear cell

CCl4 :

Carbon tetrachloride

DMEM:

Dulbecco’s modified eagle’s medium

FBS:

Fetal bovine serum

GFP:

Green fluorescent protein

HSC:

Hepatic stellate cell

IHC:

Immunohistochemistry

MMP:

Matrix metalloproteinase

mRNA:

Messenger RNA

MSC:

Mesenchymal stem cell

NOD-SCID:

Nonobese diabetic/severe combined immunodeficient

PBS:

Phosphate-buffered saline

RT-PCR:

Reverse transcription polymerase chain reaction

TNFα:

Tumor necrosis factor alpha

TGFβ:

Transforming growth factor beta

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Acknowledgment

This study was supported by Grants-in-Aid for scientific research from the Japan Society for the Promotion of Science (JSPS); Ministry of Health, Labour and Welfare, health and labour sciences research grants and Japan Science and Technology Agency (JST), the project of realization of regenerative medicine and highway. Ms. Mariko Yamada, Ms. Isako Fujimoto and Ms. Yoko Fukusumi helped us with several analyses.

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Authors and Affiliations

  1. Department of Gastroenterology and Hepatology, Yamaguchi University Graduate School of Medicine, Minami-Kogushi 1-1-1, Ube, Yamaguchi, 755-8505, Japan

    Haruko Tanimoto, Shuji Terai, Takami Taro, Yasuhiko Murata, Kouichi Fujisawa, Naoki Yamamoto & Isao Sakaida

  2. Center of Reparative Medicine, Yamaguchi University Graduate School of Medicine, Yamaguchi, Japan

    Kouichi Fujisawa

Authors
  1. Haruko Tanimoto
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  2. Shuji Terai
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  3. Takami Taro
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  4. Yasuhiko Murata
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  5. Kouichi Fujisawa
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  6. Naoki Yamamoto
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  7. Isao Sakaida
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Correspondence to Shuji Terai.

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Tanimoto, H., Terai, S., Taro, T. et al. Improvement of liver fibrosis by infusion of cultured cells derived from human bone marrow. Cell Tissue Res 354, 717–728 (2013). https://doi.org/10.1007/s00441-013-1727-2

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  • DOI: https://doi.org/10.1007/s00441-013-1727-2

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