【摘要】 本实验探讨了促性腺激素(FSH、LH)对猪卵母细胞体外成熟的影响,并最终找到了其合理的使用剂量。实验选取具有3层致密卵丘细胞层及均匀细胞质的猪卵丘-卵母细胞复合物(COCs)在体外进行成熟培养。体外成熟培养液为改良的TCM-199液,根据实验设计再添加不同浓度的FSH和LH。卵母细胞体外成熟培养条件为39℃、5%CO2与饱和湿度下,成熟培养48h。培养结束后,以第1极体排出作为卵母细胞成熟的标准。在成熟液中添加LH浓度分别为1、5、10IU/mL时,与没有添加FSH与LH的对照组(成熟率为15.9%)相比,成熟率有显著提高,前者成熟率分别为49.1%3、0.0%3、6.3%。LH浓度为1IU/mL的实验组成熟率最高,显著高于对照组(P<0.05);然后以成熟液中只添加浓度为1IU/mLLH为对照组,实验组添加FSH浓度分别为1、5、10IU/mL。结果表明,添加FSH浓度为5IU/mL和10IU/mL时其成熟率分别为40.1%、27.6%,低于对照组(49.0%)。添加FSH浓度为1IU/mL时其成熟率(49.1%)高于对照组(49.0%),但差异不显著(P>0.05)。试验表明,随着FSH与LH添加浓度逐渐增加,成熟率呈下降趋势。添加FSH与LH浓度均为1IU/mL可得到最佳的成熟效果。更多还原

【Abstract】 The objective of the present study was to examine the effect of Follicular stimulating hormone(FSH) and Luteinising hormone(LH) on in vitro maturation of porcine oocytes,and find the rational doses of them.Cumulus-oocyte complexes(COCs) with at least three layers of unexpanded cumulus cells and evenly granulated cytoplasm were selected.The basic medium for maturational culture of COCs was modified medium 199(mTCM-199) supplemented with 4.12mmol/L calcium lactate,0.91mmol/L sodium pyruvate,3.05mmol/L D-Glucose.Thereafter,mM199 was supplemented with different densities of FSH and LH(Ningbo,China).The COCs were culutured in the incubator at 39℃ in an atomosphere of 5% CO2.After culture for maturation for 48h,some of the oocytes were denuded to examine the presence of PBⅠ.The nuclear maturation were recorded as percentage of oocytes with PBⅠ.LH at 1,5,10 IU/mL,was supplemented in the maturation medium.the maturation rate of the treated groups was highly improved compared with the control(15.9%),They were 49.1%、30.0% and 36.3% separately.The highest maturation rate presented in group with LH at 1 IU/mL and was significantly different compared with the control(P<0.05).Then take the group with LH at 1 IU/mL as the control,FSH at 1,5,10 IU/mL and LH at 1 IU/mL were supplemented in the treated groups.The results showed the maturation rates of supplementation of FSH at 5 and 10 IU/mL(40.1%、27.6%) were lower than the control(49.0%),supplementation of FSH at 1 IU/mL improved the maturation rate(49.1%),but was not significantly different compared with the control(49.0%)(P>0.05).The results also showed that the maturation rate descended with the increase of the supplementation of FSH or LH.It was concluded that supplementation of FSH and LH,each at 1 IU/mL,can mostly improved the maturation rate.更多还原