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Title: EFFECTS OF STEM CELL FACTOR (SCF) AND GRANULOCYTE MACROPHAGE-COLONY STIMULATING FACTOR (GM-CSF) ON IN VITRO SURVIVAL OF PORCINE TYPE A SPERMATOGONIA.

Author
item DIRAMI, GHENIMA - GEORGETOWN UNIV MED CTR
item RAVINDRANATH, NEELAKANTA - GEORGETOWN UNIV MED CTR
item Pursel, Vernon
item DYM, MARTIN - GEORGETOWN UNIV MED CTR

Submitted to: Biology of Reproduction
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/11/1999
Publication Date: N/A
Citation: N/A

Interpretive Summary: Spermatogenesis is initiated by divisions of type A spermatogonial stem cells that reside in the testes. Little is known regarding factors that regulate this cell population. In order to obtain a better understanding of the biology of these stem cells, type A spermatogonia were isolated from the testes of 80-day-old pigs, cultured in a defined medium, and the effects of two growth factors on cell survival were studied. At low concentrations granulocyte macrophage-colony stimulating factor stimulated cell survival but at higher concentrations this growth factor inhibited survival. In contrast, stem cell factor enhanced cell survival only at high concentrations. Combining the two growth factors did not increase cell survival. These findings will be useful to scientist that are interested in maintaining long term cultures of stem cell populations for use in studies involving genetic manipulation.

Technical Abstract: 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 hr in Dulbecco's modified Eagle's medium/Hain's F12 medium (DMEM/Fl2) or a potassium-rich medium derived by the simplex optimization method (KSOM). At the end of the 120-hr culture period, 3050% 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. Combination of SCF with GMCSF had no significant effect on the percent 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.