摘要
目的：观察表皮生长因子（EGF）和干细胞因子（SCF）体外促小鼠生精细胞增殖、分化的效应。方法：对7~8日龄雄性昆明小鼠生精细胞进行混合细胞体外培养,在培养液中分别添加不同浓度（5、10、20、40、100 ng/ml）的EGF和SCF,并进行EGF和SCF的交互实验,观察生精细胞的存活率和形态学变化,并对粗线期精母细胞特异性磷蛋白基因（P19）、单倍体精子细胞特异性转化蛋白基因（TP1）及染色体倍性进行检测。结果：加入EGF或SCF 2-4 d,各组均可见不同程度的细胞增殖,细胞呈团或族状,以20 ng/ml EGF组和40 ng/ml SCF组最为明显;培养第7天,单一添加20 ng/ml EGF或40 ng/ml SCF组生精细胞数和存活率显著高于与其它各组（P〈0.05）,且40 ng/ml SCF组P19/TP1基因表达显著低于其它各组,单倍体细胞率显著高于其它各组（P〈0.05）。EGF与SCF配伍时,可显著增加体外培养后生精细胞数（P〈0.05）。结论：在混合生精细胞体外培养体系中,添加一定浓度的EGF和SCF可显著提高生精细胞数和存活率,而且SCF可提高单倍体精子的形成率;两者交互实验时,对细胞增殖有一定的叠加效应。
Objective： To study the promoting effects of the epidermal growth factor （EGF） and stem cell factor （SCF） on the proliferation and differentiation of spermatogenic eells in mice. Methods： We cocultured in vitro spermatogenic cells of male mice aged 7 -8 days in the medium with EGF and/or SCF at the concentrations orS, 10, 20, 40 and 100 ng/ml, respectively. Then we ob- served the survival rate and morphological changes of the spermatogenic cells, detected the expressions of the pachytene-specific phos- phoprotein gene （P19） and haploid sperm cell-specific transition protein gene （ TPI ）, and analyzed the ploidy of the cells. Results： After cocuhured with EGF or SCF for 2 -4 days, the spermatogenic cells began to proliferate in masses or chains in all concentration groups, most obviously in the 20 ng/ml EGF and 40 ng/ml SCF groups. At 7 days, both the number and survival rate of spermatogenic cells were significantly higher in the 20 ng/ml EGF and 40 ng/ml SCF groups than in the others （P 〈0.05）, and meanwhile, the PI9/TP1 ratio was obviously decreased and the rate of haploid sperm markedly increased in the 40 ng/ml SCF group （ P 〈0. 05 ）. Combination of EGF and SCF remarkably promoted the proliferation of the spermatogenic cells （ P 〈 0.05 ）. Conclusion ： Both EGFand SCF could increase the number and survival rate of spermatogenic cells. SCF could promote the formation of haploid sperm, and the combination of the two cytokines could enhance the effect on the proliferation of spermatogenic cells.
出处
《中华男科学杂志》
CAS
CSCD
2014年第8期679-683,共5页
National Journal of Andrology
基金
安徽省自然科学基金(090413271X)~~
