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First published online 17 August 2005
doi: 10.1242/dev.02004
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1 Horizontal Medical Research Organization, Graduate School of Medicine, Kyoto
University, Kyoto 606-8501, Japan
2 Department of Molecular Genetics Graduate School of Medicine, Kyoto
University, Kyoto 606-8501, Japan
3 RIKEN, Bioresource Center, Ibaraki 305-0074, Japan
4 Experimental Research Center for Infectious Diseases, Institute for Virus
Research, Kyoto University, Kyoto 606-8507, Japan
5 Medical Research Institute, Tokyo Medical and Dental University, Tokyo
101-0062, Japan
6 Department of Molecular and Cell Genetics, School of Life Sciences, Faculty of
Medicine, Tottori University, Yonago, Tottori 683-8503, Japan
7 Department of Pathology and Biology of Diseases, Graduate School of Medicine,
Kyoto University, Kyoto 606-8501, Japan
* Author for correspondence (e-mail: tshinoha{at}virus.kyoto-u.ac.jp)
Accepted 18 July 2005
Although stem cells are believed to divide infinitely by self-renewal
division, there is little evidence that demonstrates their infinite
replicative potential. Spermatogonial stem cells are the founder cell
population for spermatogenesis. Recently, in vitro culture of spermatogonial
stem cells was described. Spermatogonial stem cells can be expanded in vitro
in the presence of glial cell line-derived neurotrophic factor (GDNF),
maintaining the capacity to produce spermatogenesis after transplantation into
testis. Here, we examined the stability and proliferative capacity of
spermatogonial stem cells using cultured cells. Spermatogonial stem cells were
cultured over 2 years and achieved 
1085-fold expansion. Unlike
other germline cells that often acquire genetic and epigenetic changes in
vitro, spermatogonial stem cells retained the euploid karyotype and
androgenetic imprint during the 2-year experimental period, and produced
normal spermatogenesis and fertile offspring. However, the telomeres in
spermatogonial stem cells gradually shortened during culture, suggesting that
they are not immortal. Nevertheless, the remarkable stability and
proliferative potential of spermatogonial stem cells suggest that they have a
unique machinery to prevent transmission of genetic and epigenetic damages to
the offspring, and these characteristics make them an attractive target for
germline modification.
Key words: Culture, Genomic imprinting, Karyotype, Spermatogenesis, Stem cell, Telomere
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