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MBC in Press, published online ahead of print January 12, 2005
Mol. Biol. Cell 10.1091/mbc.E04-07-0652

A more recent version of this article appeared on March 1, 2005
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Submitted on July 31, 2004
Accepted on December 20, 2004

Immortalization of Human Fetal Cells: The Life Span of Umbilical-Cord-Blood-derived Cells Can Be Prolonged without Manipulating p16INK4a/RB Braking Pathway

Masanori Terai,*{dagger} Taro Uyama,* Tadashi Sugiki,* Xiao-Kang Li,{ddagger} Akihiro Umezawa,*{sect}|| and Tohru Kiyono{dagger}

Departments of *Reproductive Biology and Pathology and {ddagger}Innovative Surgery, National Research Institute for Child Health and Development, Tokyo, Japan; {dagger}Virology Division, National Cancer Center Research Institute, Tokyo, Japan; {sect}Department of Pathology, Keio University School of Medicine, Tokyo, Japan

Monitoring Editor: Lawrence Goldstein

Human umbilical-cord-blood-derived mesenchymal stem cells (UCBMSCs) are expected to serve as an excellent alternative to bone marrow-derived human mesenchymal stem cells. However, it is difficult to study them because of their limited life span. To overcome this problem, we attempted to produce a strain of UCBMSCs with a long life span and investigate whether the strain could maintain phenotypes in vitro. UCBMSCs were infected with retrovirus carrying the human telomerase reverse transcriptase (hTERT) to prolong their life span. The UCBMSCs underwent 30 population doublings (PDs) and stopped dividing at PD 37. The UCBMSCs newly established with hTERT (UCBTERTs) proliferated for >120 PDs. The p16INK4a/RB braking pathway leading to senescence can be inhibited by introduction of Bmi-1, a polycomb-group gene, and human papillomavirus type 16 E7, but the extension of the life span of the UCBMSCs with hTERT did not require inhibition of the p16INK4a/RB pathway. The characteristics of the UCBTERTs remained unchanged during the prolongation of life span. UCBTERTs provide a powerful model for further study of cellular senescence and for future application to cell-based therapy using umbilical cord blood cells.

||Corresponding author. E-mail: umezawa{at}1985.jukuin.keio.ac.jp






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