牛体外受精胚胎衍生干细胞能力影响因素的研究   总被引：1，自引：0，他引：1  

李碧春  陈国宏  吴信生  徐琪  吴圣龙  蔡中涛 《畜牧兽医学报》2006,37(9):928-932

以牛卵巢卵母细胞体外受精获取囊胚期胚胎，比较体外受精牛胚胎不同获取内细胞团的方法和不同培养液对体外培养胚胎干细胞能力的影响。结果表明，囊胚期胚胎不除去透明带而直接培养产生胚胎干细胞，初次克隆率为55％；胰酶法去透明带分离的内细胞团（ICM）培养产生胚胎干细胞，初次克隆率为90％。免疫外科法去透明带分离的ICM在4种不同的培养液中，初次克隆率分别为16．4％、11．5％、21．8％、18．2％，但是其分离的ICM经传代后形成的衍生细胞不易发生分化，经过4～6代的传代，仍然保持其完整的形态，说明免疫外科法是一种理想的牛ICM分离法，培养液为D20＋LIF（40ng／mL）可用于牛胚胎干细胞培养。  相似文献   

Estimation of the Optimal Timing of Fertilization for Embryo Development of In Vitro-Matured Bovine Oocytes Based on the Times of Nuclear Maturation and Sperm Penetration     

Keisuke KOYAMA  Sung-Sik KANG  Weiping HUANG  Yojiro YANAGAWA  Yoshiyuki TAKAHASHI  Masashi NAGANO 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2014,76(5):653-659

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激素和表皮生长因子对牛卵母细胞体外受精的影响   总被引：1，自引：1，他引：1  

魏红芳  昝林森  徐照学  辛小玲 《安徽农业科学》2007,35(33):10726-10727

[目的]研究激素和表皮生长因子(EGF)对牛卵母细胞体外受精的影响。[方法]通过对牛进行体外受精和受精卵体外培养,研究了不同血清、激素及EGF对卵母细胞体外成熟和胚胎体外发育的影响。[结果]单独添加ECS组卵母细胞的成熟率、卵裂率、桑囊率均最高,分别为81.39%5、6.05%、30.94%,且卵裂率、桑囊率与添加NBS组的差异显著。在培养液中添加EGF和激素能促进卵母细胞的成熟和卵裂的发生,其成熟率、卵裂率分别为82.50%、83.33%和61.43%、62.21%,均比未添加EGF组的高,差异显著,并且EGF与激素间有协同作用。[结论]单独添加10%ECS对卵母细胞的作用最理想,EGF对牛卵母细胞体外成熟及卵裂有良好的促进作用。  相似文献   

精子提取物对绵羊显微授精受精率及胚胎发育影响的研究     

顾文艺  Kotaras P J  Seamark R F  郭志勤 《西北农业学报》1996,5(4):1-4

将绵羊精子可溶性撮物与上不运动绵羊精人同注射于体外成熟的绵羊卵母细胞并在体外培养，结果发现，共同注射的卵母细胞卵裂率、２￣１６细胞发育率，桑椹胚发率育，囊胚发育率均显著高于对照组，其中卵母细胞卵裂率和２￣１６细胞发育率还显著高于精子单独注射组；桑椹胚，囊胚发育率明显高于精子单独注射组。  相似文献   

奶山羊精子体外获能与体外受精   总被引：1，自引：0，他引：1  

钱菊汾  郝志明  张涌 《西北农林科技大学学报(自然科学版)》1992,20(Z1)

供体用FSH+LH超排,卵母细胞从注LH后约30h回收。将获能精子与10枚卵母细胞在受精液(mDM+输卵管上皮细胞)和培养液(mDM+30％供体羊血清)中连续培养(38℃)24h后,有6枚卵受精,抛出第二极体。将6枚受精卵移植到3只受体中,有1只妊娠,产出1只试管母山羊。初步表明,本试验的奶山羊精子体外获能与体外受精程序是简便而有效的。  相似文献   

绵羊体外受精的TCM199渐变培养体系的研究     

郑小敏  史远刚  张夫千  赵鹏  马甫行  李玲玲 《吉林农业大学学报》2007,29(4):429-432

应用TCM199作卵母细胞体外成熟、体外受精及早期胚胎培养的基础培养液,再添加不同组分组成TCM199体外受精渐变培养体系,进行绵羊体外受精的研究.结果表明:TCM199体外受精渐变培养体系(试验组)与目前沿用的常规体外受精体系(对照组)相比较,虽然成熟率差异不显著(81.3%,80.4%),但能显著提高绵羊卵母细胞体外受精后的卵裂率(72.1%,50.8%)和桑囊率(94.8%,24.5%).这说明TCM199体外受精渐变培养体系能有效地克服早期胚胎发育阻断.  相似文献   

In vitro maturation and fertilization of prepubertal and pubertal black Bengal goat oocytes   总被引：1，自引：0，他引：1  

Momena Khatun  Mohammad Musharraf Uddin Bhuiyan  Jalal Uddin Ahmed  Aminul Haque  Mohammad Bozlur Rahman  Mohammed Shamsuddin 《Journal of veterinary science (Suw?n-si, Korea)》2011,12(1):75-82

Oocytes retrieval, in vitro maturation (IVM) and fertilization (IVF) efficiency are inevitable steps towards in vitro production of embryos. In the present study, these parameters were investigated in the ovaries of prepubertal (n = 31) and pubertal (n = 61) black Bengal goats obtained from a slaughterhouse. Nuclear maturation was evaluated upon aspiration and following IVM in TCM-199 (Earle''s salt with L-glutamine and sodium bicarbonate) for 27 h at 39℃ under 5% CO₂ in humidified air. The oocytes retrieval and efficiency (mean ± SD) per prepubertal and pubertal goats were 5.2 ± 0.6 and 6.8 ± 0.6, and 77.3 ± 0.1% and 80.5 ± 0.6%, respectively. Anaphase I - telophase I stages differed significantly (7.3 ± 0.8 vs. 2.6 ± 0.2, p < 0.05) between the two groups of goats. After IVM, the percentages of metaphase II were significantly higher (66.3 vs. 60.3, p < 0.05) in pubertal goats than in their prepubertal counterparts. The percentages of normal in vitro fertilization (IVF) in Fert-Tyrode''s albumin lactate pyruvate of pubertal goat oocytes did not differ between Percoll and swim-up sperm separation methods (36.7 ± 0.9% vs. 32.7 ± 1.3%, p > 0.05). Furthermore, sperm capacitation by heparin alone or in combination with ionomycin did not lead to a significant increase in the normal fertilization rate (34.8 ± 1.7 vs. 32.2 ± 1.5%, respectively) in the oocytes of pubertal goats. In conclusion, the ovaries of pubertal black Bengal goats obtained from the slaughterhouse could be used for in vitro embryo production. However, further optimization of the IVM and IVF techniques are necessary for satisfactory in vitro embryo production.  相似文献   

细胞因子对绵羊体外受精胚胎发育的影响   总被引：2，自引：0，他引：2  

雷蕾  张富春  张家新  钟喆  张大铭  郭志勤 《畜牧兽医学报》2002,33(4):344-348

绵羊卵巢采自乌鲁木齐市屠宰场，卵母细胞体外成熟24－25h后，由体外获能的附睾精子授精，50h后，将卵裂至2－8细胞的绵羊胚胎移入添加了不同浓度细胞因子的培养液中继续培养，结果表明，加入5、10、15μg/ml胰岛素（Insulin)分别得到17.46%、23.81%、22.95%的囊胚率，与对照组9.52%相比差异显著（P＜0.05)。高浓度（500μg/ml)的粒细胞巨噬细胞集落刺激因子（Granulocyte-macrophage colony stimulating factor,GM-CSF)可获得21.82%的囊胚率，与对照组7.27%相比差异显著（P＜0.05)。胚胎培养液中分别加入10、100、500ng/ml的白细胞介素－3（Interleukin-3,IL-3)、白细胞介素－8（IL－8）和神经生长因子（Nuclear growth factor,NGF)与对照组相比，囊胚率差异不显著。本研究表明，胰岛素、GM－CSF对绵羊体外受精胚胎早期发育有促进作用，IL－3、IL－8、NGF对绵羊胚胎的早期发育无明显作用。  相似文献   

Nuclear and cytoplasmic maturation of bovine oocytes cultured with dbc AMP, FSH and hCG     

P Liehman  T Greve  K P Xu 《Acta veterinaria Scandinavica》1986,27(4):566-574

The present investigation was undertaken to study the effect of addition of dbc AMP on bovine oocyte maturation and fertilization in vitro. The bovine oocytes isolated from 2–8 mm follicles were cultured for 26 h in TCM-199. The maturation rate (71.4 %) did not significantly increase after supplementation of the culture medium with dbc AMP (86.3 %.) or FSH + hCG (86.3 %). The in vitro fertilization rate of oocytes based on sperm penetration and presence of sperm tail in the ooplasm increased significantly in the dbc AMP (34.7 %) and the dbc AMP + FSH + hCG (33.9 %) treated groups when compared with untreated controls (17.9 %). However, dbc AMP treated oocytes were not able to secure the formation of male pronucleus 20 h after in vitro fertilization, while in oocytes matured in dbc AMP free medium both pronuclei were present in approximately 15 % of the penetrated oocytes. Also, the sperm head decondensation was blocked or slowed down by the dbc AMP treatment. It is concluded (1) that dbc AMP may improve the condition for the interaction of oocytes with spermatozoa, and (2) that the ooplasm of such dbc AMP treated oocytes apparently is not able to decandense the sperm head and transform it to the male pronucleus.  相似文献   

表没食子儿茶素没食子酸酯用于小鼠卵母细胞体外培养的效果观察     

李拥军  李太坤  李文婷  张光景  谷亚轻  尚利青  邹恒 《中国兽医杂志》2012,48(5):17-19

本试验以小鼠为动物模型,采用HTF和TCM-199两个基础培养体系,通过添加不同浓度的表没食子儿茶素没食子酸酯(EGCG),研究其对小鼠卵母细胞体外成熟、体外受精及其后期胚胎发育的影响。结果表明,在HTF和TCM-199体系中,培养卵丘-卵母细胞复合物(COCs)时EGCG的最佳添加量均为20μmol/L,可以显著提高卵母细胞的成熟率、受精率和胚胎发育率。过量添加EGCG则有负面的效应。综合比较,HTF体系的培养效果优于TCM-199体系。  相似文献