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Bone-marrow-derived cells cultured in serum-free medium reduce liver fibrosis and improve liver function in carbon-tetrachloride-treated cirrhotic mice

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Abstract

We have previously developed autologous bone marrow cell infusion (ABMi) therapy for liver cirrhosis patients. One problem associated with ABMi therapy is that general anesthesia is required to obtain 400 ml bone marrow fluid from liver cirrhosis patients. However, many patients with decompensated cirrhosis do not meet the criteria, because of decreased liver function or an increased bleeding tendency. To overcome these issues, our aim is to derive liver repair cells from small amounts of autologous bone marrow aspirates obtained under local anesthesia and to use these cells in liver cirrhosis patients. Here, we conducted, by using a mouse model, basic research aimed at achieving novel liver regeneration therapy. We cultured bone marrow cells aspirated from the femurs of C57 BL/6 Tg14 (act-EGFP) OsbY01 mice (green fluoresent protein [GFP]-transgenic mice). After 14 days of culture with serum-free medium (good manufacturing practice grade), the obtained spindle-shaped GFP-positive cells were injected (1×104 cells) via the caudal vein into mice with carbon tetrachloride (CCl4)-induced cirrhosis. Numerous cultured macrophages and some mesenchymal stem cells repopulated the cirrhotic liver. The results showed that serum albumin, liver fibrosis and liver function were significantly improved in the group treated with cultured bone marrow cells (P<0.01). Moreover, matrix metalloproteinase-9 expression was increased in the liver (P<0.01). Thus, infusion of bone-marrow-derived cultured cells improved liver function and liver fibrosis in mice with CCl4-induced cirrhosis.

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Abbreviations

BMC:

Bone marrow cell

GFP:

Green fluorescent protein

CCl4 :

Carbon tetrachloride

ABMi :

Autologous bone marrow cell infusion

MSC:

Mesenchymal stem cell

GMP:

Good manufacturing practice

PBS:

Phosphate-buffered saline

FITC:

Fluorescein isothiocyanate

PE:

Phycoreythrin

PI:

Propidium iodide

DAB:

Diaminobenzidine

FBS:

Fetal bovine serum

α-SMA:

α-Smooth muscle actin

MMP9:

Matrix metalloproteinase-9

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Acknowledgments

We are grateful to Ms. Mariko Yamada, Ms. Ihoko Fujimoto and Ms. Yoko Fukuzumi for assistance with cell culture, immunohistochemical analysis and animal care.

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Authors and Affiliations

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

    Takuya Iwamoto, Shuji Terai, Takuro Hisanaga, Taro Takami, Naoki Yamamoto, Shoko Watanabe & Isao Sakaida

Authors
  1. Takuya Iwamoto
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  2. Shuji Terai
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  3. Takuro Hisanaga
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  4. Taro Takami
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  5. Naoki Yamamoto
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  6. Shoko Watanabe
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  7. Isao Sakaida
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Corresponding author

Correspondence to Shuji Terai.

Additional information

This study was supported by Grants-in-Aid for scientific research from the Japan Society for the Promotion of Science (no. 22390150) and the Ministry of Health, Labour and Welfare, the Japan Science and Technology Agency (JST) and the Project of Realization of Regenerative Medicine and Highway.

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Iwamoto, T., Terai, S., Hisanaga, T. et al. Bone-marrow-derived cells cultured in serum-free medium reduce liver fibrosis and improve liver function in carbon-tetrachloride-treated cirrhotic mice. Cell Tissue Res 351, 487–495 (2013). https://doi.org/10.1007/s00441-012-1528-z

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  • DOI: https://doi.org/10.1007/s00441-012-1528-z

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