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BOR - Papers in Press, published online ahead of print January 16, 2008.
Biol Reprod 2008, 10.1095/biolreprod.107.066068
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biolreprod.107.066068v1
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Submitted October 14, 2007
Returned for revision November 4, 2007
Accepted December 7, 2007

Testis


Pluripotency of a Single Spermatogonial Stem Cell in Mice

Mito Kanatsu-Shinohara *, Jiyoung Lee , Kimiko Inoue , Narumi Ogonuki , Hiromi Miki , Shinya Toyokuni , Masahito Ikawa , Tomoyuki Nakamura , Atsuo Ogura , and Takashi Shinohara

* To whom correspondence should be addressed. E-mail: mshinoha{at}virus.kyoto-u.ac.jp.

Abstract
Although pluripotent stem cells were recently discovered in postnatal testis, attempts to analyze their developmental potential have led to conflicting claims that spermatogonial stem cells are pluripotent or that they lose spermatogenic potential after conversion into pluripotent stem cells. To examine this issue, we analyzed the developmental fate of a single spermatogonial stem cell that appeared during transfection experiments. After transfection of a neomycin-resistance gene into germilne stem (GS) cells, we obtained an embryonic stem-like, multipotent germline stem (mGS) cell line. Southern blot analysis revealed that the GS and mGS clones have the same transgene integration pattern, demonstrating their identical origin. The two lines, however, have different DNA methylation patterns. The mGS cells formed chimeras after blastocyst injection but did not produce sperm after germ cell transplantation, whereas the GS cells could only produce spermatozoa and did not differentiate into somatic cells. Interestingly, the GS cells expressed several transcription factors (Pou5f1, Sox2, Myc, and Klf4) required for reprogramming fibroblasts into a pluripotent state, suggesting that they are potentially pluripotent. Thus, our study provide evidence that a single spermatogonial stem cell can acquire pluripotentiality, but that conversion into pluripotent cell type is accompanied by loss of spermatogenic potential.

Key words: Testis • Developmental biology • Gametogenesis • Sertoli cells • Spermatogenesis


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