冻融牛始基卵泡在体外三维培养的最适藻酸盐浓度  

Investigation of the optimum concentration of sodium alginate in the three-dimensional culture of the freeze-thawed bovine primordial follicles

作　　者：贾琪[1] 王兴玲[1] 刘艳丽[1] 刘景[1] 郝大勇[1] 赵文杰[1] 

机构地区：[1]郑州大学第三附属医院,450052

出　　处：《中华生殖与避孕杂志》2017年第9期732-737,共6页Chinese Journal of Reproduction and Contraception

摘　　要：目的探讨冻融的牛始基卵泡在体外三维培养的最适藻酸钠浓度。方法玻璃化冷冻牛卵巢组织后复苏,加入Liberase DH酶消化,然后随机挑取始基卵泡,将其分别培养在二维的培养系统中(对照组,A组)和浓度分别为0.5%(B组)、1%(C组)、2%(D组)的藻酸钠三维培养系统中培养8 d,每隔1 d观察卵泡的生长情况,测量卵泡直径以及雌二醇(E_2)的分泌。结果随着培养天数增加,对照组中卵泡的完整性丧失,卵母细胞和颗粒细胞分离,而三维培养组中卵泡仍维持其完整性。体外培养8 d后,D组的卵泡平均直径为(57.7±8.5)μm,大于B组和C组(P<0.05);荧光染色后,D组的未受损卵泡率为68.4%,较B组、C组高,但仅B组、D组间有统计学差异(P<0.05);B组、C组、D组的E_2水平均随培养时间的延长而分泌增加,其中D组中的E_2水平(98.5±7.8)pg/mL最高,较B组、C组高,但仅B组、D组间有统计学差异(P<0.05)。结论藻酸钠三维培养系统较二维培养系统更适合始基卵泡的体外培养,体积浓度为2%的藻酸钠凝胶较低浓度的藻酸钠凝胶更适合始基卵泡的生长发育。Objective To investigate the optimum concentration of sodium alginate in the three-dimensional culture of the frozen-thawed bovine primordial follicles. Methods The vitrified bovine ovarian tissues were resuscitated. Liberase DH was added to digest the resuscitated ovarian tissue. Then the primordial follicles were randomly selected and cultured in a two-dimensional culture system（control group, group A） and in a three-dimensional culture system with 0.5%（group B）, 1.0%（group C）, 2.0%（group D） sodium alginate and cultured for 8 d. The growth of follicles were observed, the diameter of follicles and E2 hormone secretion were measured every other day.Results With the increase of culture time, the integrity of follicles in the control was lost, the oocytes and granules were separated, while the follicles in the three-dimensional culture group maintained its integrity. After cultured primordial follicles for 8 d in vitro, the diameter of the follicles in group D was（57.7±8.5） μm, which were higher than those in group B and group C, the difference was statistically significant（P〈0.05）. After fluorescent staining, the number of undamaged follicles in group D was 68.4%, which were higher than those in other two groups, but the difference was only statistically significant with group B（P〈0.05）. The levels of E2 in the three groups were increased with the prolongation of culture time, and the concentration of group D was（98.5±7.8） pg/mL, which were higher than those in other two groups, but the difference was only statistically significant with group B（P〈0.05）.Conclusion The three-dimensional culture system of sodium alginate was more suitable for the in vitro culture of the primordial follicles than the two-dimensional culture system. The sodium alginate gel with the concentration of2.0% was more suitable for the growth and development of the follicles.

关 键 词：始基卵泡 体外培养 藻酸钠三维培养 

分 类 号：R714.8[医药卫生—妇产科学]