Pluripotency of a single spermatogonial stem cell in mice.
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 germline stem cells, we obtained an embryonic stem-like, multipotent germline stem cell line. Southern blot analysis revealed that the germline stem and multipotent germline stem clones have the same transgene integration pattern, demonstrating their identical origin. The two lines, however, have different DNA methylation patterns. The multipotent germline stem cells formed chimeras after blastocyst injection but did not produce sperm after germ cell transplantation, whereas the germline stem cells could produce only spermatozoa and did not differentiate into somatic cells. Interestingly, the germline stem 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 provides evidence that a single spermatogonial stem cell can acquire pluripotentiality but that conversion into a pluripotent cell type is accompanied by loss of spermatogenic potential.
Kanatsu-Shinohara
Mito
M
Department of Molecular Genetics, Graduate School of Medicine, Kyoto University, Kyoto 606-8501, Japan. mshinoha@virus.kyoto-u.ac.jp
Lee
Jiyoung
J
Inoue
Kimiko
K
Ogonuki
Narumi
N
Miki
Hiromi
H
Toyokuni
Shinya
S
Ikawa
Masahito
M
Nakamura
Tomoyuki
T
Ogura
Atsuo
A
Shinohara
Takashi
T
D000818:Animals; D001755:Blastocyst / Q000737:chemistry; D015139:Blotting, Southern; D002135:Calcium-Binding Proteins / Q000235:genetics; D002454:Cell Differentiation; D002478:Cells, Cultured; D002999:Clone Cells / Q000166:cytology; D019175:DNA Methylation; D020869:Gene Expression Profiling; D005822:Genetic Vectors; D000090062:Kruppel-Like Factor 4; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D008822:Mice, Transgenic; D018832:Molecular Chaperones / Q000235:genetics; D039904:Pluripotent Stem Cells / Q000737:chemistry / Q000166:cytology*; D013093:Spermatogonia / Q000737:chemistry / Q000166:cytology*; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000737:chemistry / Q000166:cytology*; D014157:Transcription Factors / Q000235:genetics; D014162:Transfection
D002135:Calcium-Binding Proteins; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D018832:Molecular Chaperones; D014157:Transcription Factors; C087341:calmegin