体外培养时间和表皮生长因子对牦牛卵母细胞成熟及孤雌胚胎体外发育的影响

研究在成熟液中添加表皮生长因子（Epidermal growth factor,EGF）以及成熟时间对牦牛卵母细胞体外成熟及孤雌胚胎体外发育的影响,以确立牦牛卵母细胞体外成熟的最佳体系.结果表明,成熟液中添加EGF组的成熟率明显高于未添加组（P＜0.05）,并且牦牛卵母细胞的成熟率随着EGF添加的浓度增加也不断上升;随着体外培养时间的延长,成熟率逐渐增加,培养24 h后成熟率趋于稳定;胚胎体外培养液中添加EGF能显著提高孤雌胚胎的8-细胞和囊胚形成率（P＜0.05）.由此推测：在体外培养22-24 h,且添加40 μg/mL EGF最有利于牦牛卵母细胞体外成熟;胚胎培养液中添加40μg/mLEGF能显著提高牦牛孤雌胚胎的体外发育能力.     

温度对狗卵母细胞体外成熟培养的影响

以兰州土狗为实验动物,研究不同的温度对兰州土狗卵母细胞的体外成熟培养的影响,结果表明,采用不同温度以微滴(上覆灭菌矿物油)培养卵母细胞,其体外成熟率差异极显著.     

久效磷对小鼠早期胚胎体外发育的影响

采用小鼠早期胚胎的体外培养技术,研究了久效磷对小鼠着床前胚胎发育的影响。结果表明,剂量低于0.22mmol/L对小鼠早期胚胎无明显作用,剂量为0.34mmol/L时,70.6％的8细胞胚胎能发育至囊胚,剂量达0.45mmol/L,对小鼠早期胚胎有显著的毒害作用,只有5.7％可以发育至囊胚。在经久效磷处理的小鼠早期胚胎电镜照片上可以看到,细胞内网架结构保持完好,线粒体被破坏,数量变少,空泡变大、增多,细胞内不同结构对久效磷的敏感性有一定差异。     

牛卵母细胞的采集方法和卵巢贮存条件对其体外成熟的影响

比较了抽吸法和剖切法取得屠宰奶牛卵巢的可用卵母细胞数,并探讨了在卵巢运输过程中温度和时间对卵母细胞成熟率和卵裂率的影响.从屠宰场收集奶牛卵巢158个,用抽吸法抽取了96个卵巢,平均每个卵巢收集可用卵母细胞8.04枚(772枚/96卵巢);用切剖法切割了62个卵巢,平均每个卵巢收集可用卵母细胞18.89枚(1171枚/62卵巢).统计分析证明,两种采集方法获得的可用卵母细胞数差异极显著(p<0.01).将屠宰牛卵巢分别置于20℃～29℃和30℃～38℃的生理盐水中,在6h内收集到卵母细胞的成熟率和卵裂率分别为53.8%、31.7%和63.3%、34.3%,两者之间的差异均不显著(p>0.05).在30℃～38℃温度下,屠宰牛卵巢保存时间超过6h时,卵母细胞的成熟率和卵裂率显著下降,分别为36.1%和0%.与小于2h和2～6h相比差异极显著(p<0.01).     

不同种公牛精液对牛卵母细胞体外受精效果影响的研究

分别利用不同个体的海伏特、安格斯和荷斯坦种公牛精液进行体外受精，观察了不同品种不同个体种公牛精液的体外受精效果，利用五种海伏特种公牛精液进行体外受精后卵裂率为１７．６％～７４．７％，囊胚发育率为１．０％～３２．６％，不同个体间差异极显（Ｐ〈０．０１）。利用五种安格斯种公牛精液进行体外受精后卵裂率为３４．７％～８５．３％，囊胚发育率为６．２～４０．５％，不同个体间具极显差异（Ｐ〈０．０１）。利用     

绵羊卵母细胞不同采集方法对体外成熟培养的影响

为提高绵羊卵巢卵母细胞的采集效率,提供用于卵母细胞体外成熟培养和体外受精的更多优质的成熟卵母细胞,试验利用抽吸法和剖切法2种采集方法采集绵羊卵巢卵母细胞进行体外成熟培养并对其效果进行了对比研究.结果表明剖切法获得的卵子数量显著高于抽吸法(P<0.05),而所用时间则无显著差异(P>0.05);A、B级卵母细胞的比例在剖切法中显著高于抽吸法(P<0.05),抽吸法所采集的C级细胞显著高于剖切法(P<0.05);剖切法中卵丘扩散率和第一极体排出率要显著高于抽吸法(P<0.05).     

山羊胚胎的体外发育及冷冻保存研究

采用体外培养的方法，对山羊２～８细胞胚进行培养以后将体外发育至囊胚的胚胎进行冷冻保存，以便探讨山羊早期胚的体外培养方法和冷冻方法对体外发育胚冷冻效果的影响。将胚胎分别放入Ｍ１９９、Ｍ１９９＋输卵管上皮细胞和Ｍ１９９＋输卵管上皮细胞＋ＥＧＦ的培养液中进行培养，选取发育至囊胚期的胚胎分别以甘油和乙二醇为冷冻保护剂进行冷冻保存。结果体外培养胚的囊胚发育率在三个处理组中分别为１２．５％、７０．６％和７６．９％。使用Ｍ１９９处理组与添加输卵管上皮细胞、添加输卵管上皮细胞和ＥＧＦ的处理组之间具有极显著性差异（Ｐ＜０．０１）。冷冻解冻胚的形态正常率和发育率在以甘油和乙二醇为防冻剂的处理组中分别为６８．７、５８．３％和７８．６％、７２．８％。两组之间无明显差异，结果表明，采用与输卵管上皮细胞共同培养的方法能够提高山羊胚胎体外培养的发育率，以甘油和乙二醇为冷冻防冻剂均适合于山羊体外发育胚的冷冻保存，两者相比乙二醇的效果略优于甘油。     

羔山羊卵泡的发育及卵母细胞体外成熟的研究

为利用羔羊作供胚羊，采用三种超数排卵的方法，对出生后４０～１５０日龄的羔山羊进行超数排卵处理以后，从活体采集卵巢卵母细胞在体外进行了培养．研究结果表明，采用超数排卵的技术能够诱导未性成熟羔山羊的卵巢卵泡的发育，由超排后采集到的羔山羊卵泡卵母细胞经体外培养有继续发育成熟的能力．适宜的超排方法为ＦＳＨ１０ｍｇ＋ＬＨ１．５ｍｇ（肌肉注射），羔山羊适宜的超排日龄为４０～９０ｄ，卵母细胞的体外适宜培养时间为１８～２８ｈ．     

抗氧化剂对小鼠腔前卵泡及其卵母细胞体外发育的影响研究

通过使用改良的TCM199无血清培养系统,在96孔细胞培养板中,每孔放入一个腔前卵泡等条件下,探索添加物抗氧化剂维生素C对小鼠腔前卵泡卵母细胞体外生长发育的影响,旨在摸索一套适合小鼠腔前卵泡体外生长发育的培养体系.结果表明:随着浓度的增加,卵泡和卵母细胞的直径都不同程度地有所增长,其中50μg/m L浓度组在卵泡培养相同时间下,卵泡和卵母细胞的增长值相对于其他浓度组略大些,但各浓度组之间无论是卵泡直径增长值还是卵母细胞直径增长值都不存在显著性差异(P0.05).培养到第6天时,50和100μg/m L浓度组卵泡存活率最高,为59.5%,且50μg/m L浓度组第一极体排出率最高,为2.38%,但发生生发泡破裂率次于150μg/m L浓度组.可得结论:在改良的TCM199无血清培养系统中,添加50μg/m L维生素C有利于腔前卵泡卵母细胞的生长,而添加150μg/m L维生素C有利于生发泡破裂.     

体外注射激素对中国对虾卵巢发育的影响

研究了4种不同激素(17α-羟孕酮、5-羟色胺、前列腺素和保幼激素-Ⅲ)的体外注射对中国对虾(Penaeuschinensis)卵巢发育的作用效果．试验结果显示用100ng／g浓度的1717α-羟孕酮和5-羟色胺注射亲虾对对虾卵巢的发育有明显的刺激作用,生殖指数分别比对照组增加了50．0％和44．7％．而同样浓度的前列腺素和JH-Ⅲ的体外注射作用不明显．在相同条件下,去眼柄对对虾卵巢的发育作用最明显,生殖指数比对照组增加了150％．文中还讨论分析了几种激素的属性和对虾卵巢发育的作用机理．     

Mitochondrial functions on oocytes and preimplantation embryos

Oocyte quality has long been considered as a main limiting factor for in vitro fertilization (IVF). In the past decade, extensive observations demonstrated that the mitochondrion plays a vital role in the oocyte cytoplasm, for it can provide adenosine triphosphate (ATP) for fertilization and preimplantation embryo development and also act as stores of intracellular calcium and proapoptotic factors. During the oocyte maturation, mitochondria are characterized by distinct changes of their distribution pattern from being homogeneous to heterogeneous, which is correlated with the cumulus apoptosis. Oocyte quality decreases with the increasing maternal age. Recent studies have shown that low quality oocytes have some age-related dysfunctions, which include the decrease in mitochondrial membrane potential, increase of mitochondrial DNA (mtDNA) damages, chromosomal aneuploidies, the incidence of apoptosis, and changes in mitochondrial gene expression. All these dysfunctions may cause a high level of developmental retardation and arrest of preimplantation embryos. It has been suggested that these mitochondrial changes may arise from excessive reactive oxygen species (ROS) that is closely associated with the oxidative energy production or calcium overload, which may trigger permeability transition pore opening and subsequent apoptosis. Therefore, mitochondria can be seen as signs for oocyte quality evaluation, and it is possible that the oocyte quality can be improved by enhancing the physical function of mitochondria. Here we reviewed recent advances in mitochondrial functions on oocytes.     

牦牛胎盘活性因子生物学效应的研究

通过在培养基中加入不同浓度的牦牛胎盘粉饲养果蝇,研究牦牛胎盘活性因子对果蝇繁殖力和体重的生物学效应.结果表明:①牦牛胎盘活性因子对提高果蝇的繁殖力有显著作用,当牦牛胎盘粉浓度为0.6%时效果最显著;②牦牛胎盘活性因子对果蝇体重有影响.培养在牦牛胎盘粉浓度为2.4%培养基中的雄果蝇体重与对照组有显著差异,而培养在浓度为0.2%培养基中的雌果蝇体重与浓度为0.6%、1.2%、2.4%培养基中的雌果蝇体重有显著差异.     

Seasonal influence on the development of parthenotes and cloned embryos in pigs

Seasonal influence on the developmental ability of porcine embryos was studied by comparisons of the number of cumulus-oocytes-complexes (COCs) per ovary pair and meiotic maturation ability of oocytes in different seasons; and by the observations on developmental competence of the parthenogenetic oocytes and cloned embryos collected from different seasons. We found that the number of COCs per ovary pairs was significantly higher in spring compared to the other seasons (P<0.05). However, no significant difference was found in the rate of oocytes progressing to MⅡ in four seasons (P<0.05). In addition, the rate of blastocyst formation of parthenogenetic embryos in summer was obviously declined compared to the other seasons (P<0.05), and an increased blastocyst rate of cloned porcine embryos was found in spring compared to autumn and winter (P<0.05). The results suggest that there should be a seasonal influence on the developmental competence of porcine embryos.     

Seasonal influence on the development of parthenotes and cloned embryos in pigs

Seasonal influence on the developmental ability of porcine embryos was studied by comparisons of the number of cumulus-oocytes-complexes (COCs) per ovary pair and meiotic maturation ability of oocytes in different seasons; and by the observations on developmental competence of the parthenogenetic oocytes and cloned embryos collected from different seasons. We found that the number of COCs per ovary pairs was significantly higher in spring compared to the other seasons (P<0.05). However, no significant difference was found in the rate of oocytes progressing to MⅡ in four seasons (P<0.05). In addition, the rate of blastocyst formation of parthenogenetic embryos in summer was obviously declined compared to the other seasons (P<0.05), and an increased blastocyst rate of cloned porcine embryos was found in spring compared to autumn and winter (P<0.05). The results suggest that there should be a seasonal influence on the developmental competence of porcine embryos.     

女性年龄和获卵数对PESA-ICSI治疗结果的影响

目的:卵胞质内单精子注射(ICSI)治疗60例阻塞性无精子症患者,分析女方年龄和获卵数对ICSI结局的影响.方法:经皮附睾精子抽吸术(PESA)获得精子,女方进行常规超排卵.分析女方不同年龄组(＜30岁组、30～35岁组和＞35岁组)和不同获卵数组(1～10个组、10～19个组和＞19个组)患者行ICSI后的受精率、卵...