Can the life span of human marrow stromal cells be prolonged by bmi-1, E6, E7, and/or telomerase without affecting cardiomyogenic differentiation?

Authors

  • Yukiji Takeda,

    1. Department of Reproductive Biology and Pathology, National Research Institute for Child Health and Development, Tokyo, Japan
    2. Department of Pathology, Keio University School of Medicine, Tokyo, Japan
    3. Department of General Medicine and Clinical Investigation, Nara Medical University, Nara, Japan
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  • Taisuke Mori,

    1. Department of Reproductive Biology and Pathology, National Research Institute for Child Health and Development, Tokyo, Japan
    2. Department of Pathology, Keio University School of Medicine, Tokyo, Japan
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  • Hideaki Imabayashi,

    1. Department of Reproductive Biology and Pathology, National Research Institute for Child Health and Development, Tokyo, Japan
    2. Department of Orthopedics, Keio University School of Medicine, Tokyo, Japan
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  • Tohru Kiyono,

    1. Virology Division, National Cancer Center Research Institute, Tokyo, Japan
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  • Satoshi Gojo,

    1. Department of Cardiovascular Surgery, Saitama Medical Center, Kawagoe, Japan
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  • Shunichirou Miyoshi,

    1. Cardiopulmonary Division, Department of Internal Medicine, Keio University School of Medicine, Tokyo, Japan
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  • Naoko Hida,

    1. Department of Reproductive Biology and Pathology, National Research Institute for Child Health and Development, Tokyo, Japan
    2. Department of Pathology, Keio University School of Medicine, Tokyo, Japan
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  • Makoto Ita,

    1. Pharmacia-Keio Research Laboratories, Shinanomachi Research Park, Keio University School of Medicine, Tokyo, Japan
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  • Kaoru Segawa,

    1. Department of Virology, Keio University School of Medicine, Tokyo, Japan
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  • Satoshi Ogawa,

    1. Cardiopulmonary Division, Department of Internal Medicine, Keio University School of Medicine, Tokyo, Japan
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  • Michiie Sakamoto,

    1. Department of Pathology, Keio University School of Medicine, Tokyo, Japan
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  • Shinobu Nakamura,

    1. Department of General Medicine and Clinical Investigation, Nara Medical University, Nara, Japan
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  • Akihiro Umezawa

    Corresponding author
    1. Department of Reproductive Biology and Pathology, National Research Institute for Child Health and Development, Tokyo, Japan
    • Department of Reproductive Biology and Pathology, National Research Institute for Child Health and Development, Okura, Setagaya, Tokyo, 157-8535 Japan.
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Abstract

Background

Cell transplantation has recently been challenged to improve cardiac function of severe heart failure. Human mesenchymal stem cells (hMSCs) are multipotent cells that can be isolated from adult marrow stroma, but because of their limited life span, it is difficult to study them further. To overcome this problem, we attempted to prolong the life span of hMSCs and investigate whether the hMSCs modified with cell-cycle-associated genes can differentiate into cardiomyocytes in vitro.

Methods

We attempted to prolong the life span of hMSCs by infecting retrovirus encoding bmi-1, human papillomavirus E6 and E7, and/or human telomerase reverse transcriptase genes. To determine whether the hMSCs with an extended life span could differentiate into cardiomyocytes, 5-azacytidine-treated hMSCs were co-cultured with fetal cardiomyocytes in vitro.

Result

The established hMSCs proliferated over 150 population doublings. On day 3 of co-cultivation, the hMSCs became elongated, like myotubes, began spontaneously beating, and acquired automaticity. Their rhythm clearly differed from that of the surrounding fetal mouse cardiomyocytes. The number of beating cardiomyocytes increased until 3 weeks. hMSCs clearly exhibited differentiated cardiomyocyte phenotypes in vitro as revealed by immunocytochemistry, RT-PCR, and action potential recording.

Conclusions

The life span of hMSCs was prolonged without interfering with cardiomyogenic differentiation. hMSCs with an extended life span can be used to produce a good experimental model of cardiac cell transplantation and may serve as a highly useful cell source for cardiomyocytic transplantation. Copyright © 2004 John Wiley & Sons, Ltd.

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