In Vitro Ectopic Behavior of Porcine Spermatogonial Germ Cells and Testicular Somatic Cells.
Embryonic body-like colony formation is a unique pattern in male germ cell cultures, including spermatogonial stem cells. However, detailed information of the colony formation has not yet been sufficiently reported in male germ cell culture. To elucidate the formation of germ cell-derived colony (GDC), glial cell-derived neurotrophic factor receptor alpha-1 (GFRα-1)-positive pig germ cells were isolated using an immunomagnetic cell isolation method and labeled with red- or green-fluorescent dye. In GDC culture, red-fluorescent-labeled germ cells were evenly distributed in the wells from day 1 to 4, and they clustered together at the time of GDC formation on day 6. Interestingly, feeder cells migrated to the site of colony formation as spermatogonia carriers. Furthermore, when freshly prepared green-labeled GFRα-1-positive germ cells were added, mixed-fluorescent dye (red and green) colonies were observed. On bromodeoxyuridine (BrdU) treatment, 58% ± 3.13% of germ cells were positive to protein gene product 9.5 but negative to BrdU cells. Immunocytochemistry and reverse transcription-polymerase chain reaction results showed that cultured GDC cells were positive to stem cell- and pig germ cell-specific marker genes. In conclusion, in vitro formation of GDCs is mainly dependent on the aggregation of single germ cells as well as on the slow proliferation of germ cells.
1 Department of Stem Cell and Regenerative Biology, College of Animal Bioscience & Technology, Konkuk University , Seoul, Korea.
Kyung Hoon
KH
Lee
2 Department of Food Bioscience, College of Biomedical & Health Science, Konkuk University , Chung-ju, Korea.
Won Young
WY
Lee
1 Department of Stem Cell and Regenerative Biology, College of Animal Bioscience & Technology, Konkuk University , Seoul, Korea.
Jung Tae
JT
Do
1 Department of Stem Cell and Regenerative Biology, College of Animal Bioscience & Technology, Konkuk University , Seoul, Korea.
Chan Kyu
CK
Park
3 Department of Animal Science, College of Agriculture, Chungbuk National University , Choung-ju, Korea.
Nam Hyung
NH
Kim
1 Department of Stem Cell and Regenerative Biology, College of Animal Bioscience & Technology, Konkuk University , Seoul, Korea.
Jin Hoi
JH
Kim
4 Animal Biotechnology Division, National Institute of Animal Science , RDA, Wanju, Korea.
Hak Jae
HJ
Chung
4 Animal Biotechnology Division, National Institute of Animal Science , RDA, Wanju, Korea.
Dong Woon
DW
Kim
1 Department of Stem Cell and Regenerative Biology, College of Animal Bioscience & Technology, Konkuk University , Seoul, Korea.
Hyuk
H
Song
D000818:Animals; D000831:Animals, Newborn; D002470:Cell Survival; D002478:Cells, Cultured; D066298:In Vitro Techniques; D008297:Male; D013093:Spermatogonia / Q000166:cytology / Q000502:physiology*; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*; D013552:Swine; D013737:Testis / Q000166:cytology / Q000502:physiology*