Effects of stem cell factor and granulocyte macrophage-colony stimulating factor on survival of porcine type A spermatogonia cultured in KSOM.
Spermatogenesis is initiated with the divisions of the type A spermatogonial stem cells; however, the regulation of this stem cell population remains unknown. In order to obtain a better understanding of the biology of these cells, type A spermatogonia were isolated from 80-day-old pig testes by sedimentation velocity at unit gravity. The cells were cultured for up to 120 h in Dulbecco's modified Eagle's medium/Ham's F-12 medium (DMEM/F12) or a potassium-rich medium derived by the simplex optimization method (KSOM). At the end of the 120-h culture period, 30-50% of the spermatogonia were viable in KSOM, whereas in DMEM/F12 very few cells survived. Using KSOM as the culture medium, the effects of stem cell factor (SCF) and granulocyte macrophage-colony stimulating factor (GM-CSF) were studied. SCF significantly enhanced the percentage of cell survival at 100 ng/ml but not at lower concentrations. In comparison, GM-CSF promoted survival at relatively low concentrations (0.01, 0.1, and 1 ng/ml). At a higher dose (10 ng/ml), a significant reduction in percentage of cell survival was observed. The combination of SCF with GM-CSF had no significant effect on the percentage survival of type A spermatogonial cells. These data indicate that SCF and GM-CSF play a role in the regulation of survival and/or proliferation of type A spermatogonia.
Dirami
G
G
Department of Cell Biology, Georgetown University Medical Center, Washington, District of Columbia 20007, USA.
Ravindranath
N
N
Pursel
V
V
Dym
M
M
D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D003470:Culture Media; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology*; D007700:Kinetics; D008297:Male; D011188:Potassium / Q000494:pharmacology; D011994:Recombinant Proteins / Q000494:pharmacology; D013093:Spermatogonia / Q000502:physiology*; D019089:Stem Cell Factor / Q000494:pharmacology*; D013552:Swine
D003470:Culture Media; D011994:Recombinant Proteins; D019089:Stem Cell Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D011188:Potassium