Effects of stem cell factor and granulocyte macrophage-colony stimulating factor on survival of porcine type A spermatogonia cultured in KSOM
Effects of stem cell factor and granulocyte macrophage-colony stimulating factor on survival of porcine type A spermatogonia cultured in KSOM
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DOI：
10.1095/biolreprod61.1.225
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发表时间：
1999-07-01
影响因子：
3.6
通讯作者：
Dym, M
中科院分区：
文献类型：
--
作者：
Dirami, G;Ravindranath, N;Dym, M
精子发生始于A型精原干细胞的分裂；然而，这种干细胞群的调控仍然是未知的。为了更好地了解这些细胞的生物学特性，采用单位重力沉降速度法从80日龄猪睾丸中分离出a型精原细胞。细胞在Dulbecco's modified Eagle's medium/Ham's F-12 medium （DMEM/F12）或通过单纯形优化法（KSOM）获得的富钾培养基中培养120 h。在120 h的培养期结束时，KSOM中30-50%的精原细胞存活，而DMEM/F12中很少有细胞存活。以KSOM为培养基，研究干细胞因子（SCF）和粒细胞巨噬细胞集落刺激因子（GM-CSF）的作用。SCF在100 ng/ml浓度下显著提高细胞存活率，而在较低浓度下无显著提高。相比之下，CM-CSF在相对较低浓度（0.01、0.1和1 ng/ml）下可促进存活。在较高剂量（10 ng/ml）下，观察到细胞存活率显著降低。SCF与CM-CSF联合使用对A型精原细胞存活率无显著影响。这些数据表明，SCF和GM-CSF在调节a型精原细胞的存活和/或增殖中发挥作用。
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, CM-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 CM-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.