干细胞因子和白血病抑制因子对冻融后人原始卵泡体外生长发育的研究

Study on the Effects of Different Components Added to the Primordial Follicle Culture Medium in vitro

目的:探讨干细胞因子(stem cell factor,SCF,又称KIT配体)和白血病抑制因子(leukaemiainhibitory factor,LIF)对冻融后人原始卵泡体外培养生长发育的影响。方法:收集18例卵巢良性肿瘤手术患者的正常卵巢皮质,采用直接覆盖玻璃化冷冻法保存卵巢组织,冻融后分离卵泡,随机分成5组:对照组(A组)、10 ng/ml SCF组(B组)、10 ng/ml LIF组(C组)、10 ng/ml SCF+10 ng/ml LIF组(D组)和100 ng/ml SCF+10 ng/ml LIF组(E组),行体外培养,比较SCF和/或LIF对人原始卵泡生长发育的影响。结果:①随培养天数的增加,对照组卵泡直径增长缓慢,明显小于各实验组(P<0.05);D、E组中卵泡直径增长迅速,较B、C组差异有统计学意义(P<0.05)。②随着培养天数的增加,各组卵泡均能分泌激素。4 d后对照组卵泡激素水平增长缓慢,明显低于各实验组(P<0.05),而D、E组中卵泡激素分泌增长迅速,较B、C组差异有统计学意义(P<0.05),D、E组中卵泡激素增长速度相差不大,但在第4日差异有统计学意义(P<0.05)。结论:在体外培养液中添加SCF或LIF以及两者联合添加有利于促进卵泡的生长发育,提高E2分泌功能,且SCF和LIF联合其作用更加明显。

Objective：To explore the effects of stem cell factor（SCF） and leukaemia inhibitory factor（LIF） on human primordial follicles in vitro culture.Methods： A total of 18 ovarian tissue examples were collected and preserved by direct cover vitrification cryoapplication.Isolated follicles were divided into five groups randomly,which were control group（group A）,10 ng/ml SCF group（group B）,10 ng/ml LIF group（group C）,10 ng/ml SCF＋10 ng/ml LIF group（group D）,100 ng/ml SCF＋10 ng/ml LIF group（group E）,respectively,to compare the effects of SCF and/or LIF on human primordial follicles in vitro culture.Results： 1） The follicular diameter in group A increased slowly,and was obviously smaller than that in the experimental groups（P〈0.05）.The follicular diameter in groups D and E increased quickly,and there was a significant difference compared with groups B and C（P〈0.05）.2） Follicles in all groups had a function of hormone secretion after several days cultured in vitro.After 4 d,the level of estradiol in group A was increased slowly,and was lower than that in the experimental groups（P〈0.05）.The level of estradiol in group D or E was increased quickly,and was higher than that in group B or C（P〈0.05）.There was no difference with different density of SCF between groups D and E,but,there was a significant difference on the 4th day（P〈0.05）.Conclusion： SCF,LIF and combination of the two added to culture medium in vitro can promote follicular growth and development,raise the level of estradiol secretion,moreover,the joint action of SCF and LIF are more manifest.