两种玻璃化冷冻方法在大鼠卵巢组织冷冻中的应用效果观察  

Comparison of Effects of Two Kinds of Vitrification Methods in Freezing Ovarian Tissues in Rats

作　　者：李冬秀[1] 耿杨柳[1] 吴小华 于晓惠[1] 张敏[1] 徐清华[1] 许鹏宇[1] 

机构地区：[1]解放军白求恩国际和平医院妇产科生殖医学中心,石家庄050082

出　　处：《解放军医药杂志》2015年第5期9-12,共4页Medical & Pharmaceutical Journal of Chinese People’s Liberation Army

基　　金：河北省计划生育委员会科研基金项目(2012-C02)

摘　　要：目的比较直接覆盖玻璃化冷冻(direct cover vitrification,DCV)及常规玻璃化冷冻(conventional vitrification,CV)在大鼠卵巢组织冷冻中的应用效果。方法将雌性Wistar大鼠30只随机分为A、B、C组,每组10只,切除大鼠双侧卵巢组织。A组为对照组,B组和C组分别应用CV和DCV冻存2周。复苏后观察3组卵泡的形态,并用末端脱氧核酰转移酶介导的脱氧尿苷三磷酸标记实验(TUNEL)检测各组卵泡的凋亡率。结果 A组各级卵泡形态正常;B组卵泡形状不规则,卵母细胞核固缩;C组较大的窦状卵泡损伤。除C组原始卵泡外,B组和C组形态正常的各级卵泡比例均低于A组(P<0.05);C组形态正常的各级卵泡比例高于B组(P<0.05)。B、C组卵泡凋亡率高于A组,且C组低于B组(P<0.01)。结论 DCV较CV卵巢组织效果更好,可以有效保存卵巢组织中的原始卵泡。Objective To compare the effects of direct cover vitrification（DCV） and conventional vitrification（CV） in freezing ovarian tissues in rats. Methods A total of 30 mature female Wistar rats were randomly divided into group A,B and C（ n = 10 for each group）,and bilateral ovarian tissues of all rats were cut off. Group A was the control group; group B and C were respectively freezed using CV or DCV method for 2 weeks. After thawed,in the three groups,the follicular appearances were observed,and follicular apoptosis rates were detected using TUNEL（ terminal deoxynucleotidyl transferase-mediated d UTP nick end labeling） method. Results The appearances of each grade were normal in group A; the follicular appearances were irregular and oocyte karyopyknosis was found in group B; the bigger sinusoid ovarian follicles were damaged in group C. The normal rates of follicular appearances in all grades of group B and C were significantly lower than those in group A except for that of primordial follicles in group C（ P〈0. 05）; the normal ratios of follicular appearances in all grades of group B were significantly higher than those in group B（ P〈0. 05）. The follicular apoptosis rates in group B and C were significantly higher than that in group A,and the rate in group C was significantly lower than that in group B（ P〈0. 01）. Conclusion Primordial follicles in the ovarian tissues can be better preserved by DCV than by CV.

关 键 词：卵巢 玻璃化作用 低温保存 大鼠 Wistar 

分 类 号：R715[医药卫生—妇产科学] R-332[医药卫生—临床医学]