玻璃化冷冻对人类卵巢皮质组织形态学和凋亡的影响  

Effect of Vitrification on Morphology and Apoptosis of Human Ovarian Cortex Tissue

作　　者：汪翔[1] 方丛[1] 狄春光 史亚男[1] 梁晓燕[1] 

机构地区：[1]中山大学附属第六医院生殖医学研究中心,广东广州510655

出　　处：《中山大学学报（医学科学版）》2015年第4期545-550,共6页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(81070495);广东省自然科学基金(S2013010013404)

摘　　要：【目的】检测玻璃化冷冻方法对人卵巢皮质组织形态学和凋亡情况的影响,为其进一步应用于肿瘤患者卵巢生育力保存的安全性及可行性奠定基础。【方法】卵巢组织来自4例因疾病原因行部分卵巢切除术的患者,术后病理均提示无肿瘤细胞转移或其他干扰实验结果的病灶存在。将获得的卵巢组织分离出皮质切割成薄片,采用自身对照,随机平均分为新鲜对照组(F组)和玻璃化冷冻组(V组),两组薄片进行HE染色后,比较两组卵巢组织冻融前后的卵泡形态与各级卵泡所占比例及组织间质形态,原位末端转移酶标记(TUNEL)技术和Westernblot检测卵巢组织冻融前后细胞的凋亡情况。【结果】F组的原始卵泡、初级卵泡、次级卵泡分别为(55.5±4.9)%、(34.6±1.6)%和(9.9±7.6)%,V组的各级卵泡比例分别为(51.2±5.1)%、(36.7±1.6)%和(12.1±3.5)%,两组间比较差异无统计学意义(P>0.05);使用TUNEL法比较两组的组织内细胞凋亡情况,其差异无统计学意义(P>0.05);进一步使用Western Blot法检测两组组织内Cleaved Caspase-3的表达量,其结果与TUNEL法一致,两组间差异无统计学意义(P>0.05)。【结论】玻璃化冷冻法对卵巢组织的形态学和凋亡没有明显的影响,但其大规模临床应用仍有待更多的研究论证。【Objective】 To examine the effect of vitrification on morphology and apoptosis in human ovarian cortex tissue.【Methods】 Ovarian tissues came from 4 cases of partial ovariectomy, with necessity of diagnosis or treatment. In each case, the cortex was cut into small pieces. After that they were randomly divided into Fresh Group and Vitrification Group. The morphology of follicles and stromal cells were examined by HE staining, and the cell apoptosis in situ was analyzed by TUNEL assay. Furthermore, we quantified the expression of Cleaved Caspase-3（an indicator of apoptosis）. 【Results】 The percentage of primordial follicles, primary follicles and secondary follicles in Fresh Group are（55.5 ± 4.9）%,（34.6 ± 1.6）%, and（9.9 ± 7.6）%, respectively, while the percentage in Vitrification Group are（51.2 ± 5.1）%,（36.7 ± 1.6）%, and（12.1 ± 3.5）%. The percentage of primordial follicles,primary follicles and secondary follicles and the apoptosis in situ show no significance between these two groups. Besides, the expression of Cleaved Caspase-3 in ovarian tissue is the same between two groups. 【Conclusion】 The strategy of vitrification of human ovarian tissue is viable, and still needs more researches.

关 键 词：玻璃化冷冻 人卵巢组织 形态学变化 细胞凋亡 

分 类 号：R711.6[医药卫生—妇产科学]