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

Yukiji Takeda, Taisuke Mori, Hideaki Imabayashi, Tohru Kiyono, Satoshi Gojo, Shunichirou Miyoshi, Naoko Hida, Makoto Ita, Kaoru Segawa, Satoshi Ogawa, Michiie Sakamoto, Shinobu Nakamura, Akihiro Umezawa

Research output: Contribution to journalArticle

78 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)833-845
Number of pages13
JournalJournal of Gene Medicine
Volume6
Issue number8
DOIs
Publication statusPublished - 2004 Aug

Fingerprint

Telomerase
Stromal Cells
Mesenchymal Stromal Cells
Bone Marrow
Cardiac Myocytes
Cell Transplantation
Azacitidine
cdc Genes
Skeletal Muscle Fibers
Retroviridae
Heart Transplantation
Action Potentials
Theoretical Models
Heart Failure
Transplantation
Immunohistochemistry

Keywords

  • Bmi-1
  • Cardiomyocytes
  • Immortalization
  • Marrow stroma
  • Papillomavirus
  • Senescence

ASJC Scopus subject areas

  • Genetics

Cite this

Can the life span of human marrow stromal cells be prolonged by bmi-1, E6, E7, and/or telomerase without affecting cardiomyogenic differentiation? / Takeda, Yukiji; Mori, Taisuke; Imabayashi, Hideaki; Kiyono, Tohru; Gojo, Satoshi; Miyoshi, Shunichirou; Hida, Naoko; Ita, Makoto; Segawa, Kaoru; Ogawa, Satoshi; Sakamoto, Michiie; Nakamura, Shinobu; Umezawa, Akihiro.

In: Journal of Gene Medicine, Vol. 6, No. 8, 08.2004, p. 833-845.

Research output: Contribution to journalArticle

Takeda, Y, Mori, T, Imabayashi, H, Kiyono, T, Gojo, S, Miyoshi, S, Hida, N, Ita, M, Segawa, K, Ogawa, S, Sakamoto, M, Nakamura, S & Umezawa, A 2004, 'Can the life span of human marrow stromal cells be prolonged by bmi-1, E6, E7, and/or telomerase without affecting cardiomyogenic differentiation?', Journal of Gene Medicine, vol. 6, no. 8, pp. 833-845. https://doi.org/10.1002/jgm.583
Takeda, Yukiji ; Mori, Taisuke ; Imabayashi, Hideaki ; Kiyono, Tohru ; Gojo, Satoshi ; Miyoshi, Shunichirou ; Hida, Naoko ; Ita, Makoto ; Segawa, Kaoru ; Ogawa, Satoshi ; Sakamoto, Michiie ; Nakamura, Shinobu ; Umezawa, Akihiro. / Can the life span of human marrow stromal cells be prolonged by bmi-1, E6, E7, and/or telomerase without affecting cardiomyogenic differentiation?. In: Journal of Gene Medicine. 2004 ; Vol. 6, No. 8. pp. 833-845.
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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.",
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AU - Mori, Taisuke

AU - Imabayashi, Hideaki

AU - Kiyono, Tohru

AU - Gojo, Satoshi

AU - Miyoshi, Shunichirou

AU - Hida, Naoko

AU - Ita, Makoto

AU - Segawa, Kaoru

AU - Ogawa, Satoshi

AU - Sakamoto, Michiie

AU - Nakamura, Shinobu

AU - Umezawa, Akihiro

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N2 - 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.

AB - 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.

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KW - Papillomavirus

KW - Senescence

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