25-羟基胆甾醇对鸡原始生殖细胞增殖的影响

Effect of 25-hydroxycholesterol on Proliferation of Chicken Primordial Germ Cells

本研究为了优化现有的鸡PGCs(原始生殖细胞)的培养系统,为今后研究生殖细胞发育与生产转基因鸡奠定良好基础。分别用含0μmol/L(对照组)、0.1μmol/L、0.5μmol/L、1.0μmol/L 25-羟基胆甾醇的培养液培养鸡PGCs,5 d后细胞计数检测其细胞增殖情况。与对照组(不添加25-羟基胆甾醇的鸡PGCs培养液)相比,0.1μmol/L处理组细胞数目没有显著差异,0.5μmol/L和1μmol/L处理组细胞数目均显著增加。对1.0μmol/L 25-羟基胆甾醇处理的鸡PGCs用RT-PCR、细胞免疫染色、细胞注射迁移的方法检测处理后的生物学特性,发现处理后的鸡PGCs仍然保持正常的生殖细胞生物学特性。本研究表明25-羟基胆甾醇能有效的促进鸡P GCs的增殖,且不改变鸡PGCs的生物学特性。

The objective of this study is to optimize the culture system of chicken PGCs （primordial germ cells）, and thereby facilitate future research on germ cell development and production of transgenic chickens. Chicken PGCs were treated with 25-hydroxycholesterol （25-OHC） at 0 μmol/L （control group）, 0.1 μmol/L, 0.5 μmol/L and 1.0 μmol/L for 5 d, respectively. The proliferation of chicken PGCs was examined through cell counting. Compared with the control group, the cell number in 0.1 μmol/L treated group was not significantly different, but the cell number in 0.5 μmol/L and 1.0 μmol/L treated group was significantly increased. RT-PCR, immtmocytochemistry and migration assay demonstrated that biological characteristics of the chicken PGCs after treated with 1.0 μmol/L 25-OHC were similar to the untreated group. This paper demonstrated 25-OHC could effectively promote the proliferation of chicken PGCs without altering the biological characteristics of chicken PGCs.