通过优化超排后合笼时间高效获取小鼠孵化囊胚

doi:10.16431/j.cnki.1671-7236.2015.05.033

›› 2015, Vol. 42 ›› Issue (5): 1253-1258.doi: 10.16431/j.cnki.1671-7236.2015.05.033

通过优化超排后合笼时间高效获取小鼠孵化囊胚

翟椿东, 顾美超, 刘迪, 王力红, 齐晓龙, 张嘉楠, 刘云海, 倪和民, 郭勇

北京农学院动物科学技术学院, 北京 102206

修回日期:2015-03-04 出版日期:2015-05-20 发布日期:2015-06-02
通讯作者: 郭勇 E-mail:y63guo@126.com
作者简介:翟椿东(1986-),男,山东潍坊人,硕士生,研究方向:动物产科及胚胎工程,E-mail:15300036918@163.com
基金资助:
2013年度国家自然基金面上项目:牛体内外植入前胚胎IFNτ的差异表达及对子宫上皮细胞ISG15和Wnt7a表达的体外诱导 (31272526);2013年度北京市教委北京市属高等学校创新团队建设与教师职业发展计划项目:体细胞转基因克隆肉牛新品系培育与利用(PXM2013_014207_000067)

Study on Time Period Model for Efficiently Collecting Hatched Blastocysts Flushed from Murine Uterus

ZHAI Chun-dong, GU Mei-chao, LIU Di, WANG Li-hong, QI Xiao-long, ZHANG Jia-nan, LIU Yun-hai, NI He-min, GUO Yong

College of Animal Science and Technology, Beijing University of Agriculture, Beijing 102206, China

Revised:2015-03-04 Online:2015-05-20 Published:2015-06-02

摘要/Abstract

摘要： 本试验旨在优化小鼠超排后的合笼时间,高效获取小鼠孵化囊胚。试验选用150只ICR系8周龄雌性小鼠,随机分为5组,同一时间超排处理后雌、雄按1:1于18:00、19:00、20:00、21:00合笼过夜,次日上午08:00查栓,发现阴道栓这为妊娠第1天(D1)。取妊娠第5天(D5)小鼠处死,剪取双侧子宫角,冲取胚胎。统计每组冲取胚胎的总数及孵化囊胚/未孵化囊胚的比值,作为胚胎获取效率的评价指标;统计内细胞团数/滋养外胚层细胞团数,作为评价胚胎质量的参考指标。结果发现,在数量上组Ⅰ、组Ⅱ、组Ⅲ囊胚数差异不显著(P>0.05),但有增高趋势,组Ⅳ囊胚数显著高于其他3组(P<0.05)。组Ⅰ、组Ⅱ、组Ⅲ、组Ⅳ内细胞团数/滋养外胚层细胞团数分别为23.18%、23.55%、21.72%和23.28%,各组间差异不显著(P>0.05)。结果表明,组Ⅳ所对应的合笼时间获取小鼠孵化囊胚获取效率最高,胚胎囊胚质量无明显差异。

关键词: 小鼠; 孵化囊胚; 超数排卵; 合笼时间

Abstract: The persent study was aimed to investigate the optimization of mice cage time after superovulation, and efficient acquisition of mouse hatched blastocysts.The experiment selected 150 ICR female mice aged 8 weeks, were randomly divided into 5 groups, the same time superovulation treatment according to 1:1, after the male and female in 18:00, 19:00, 20:00, 21:00 alloy cage overnight, the next day as early as 08:00 check, found that vaginal suppository for the first day of the pregnancy (D1).Take the pregnant the fifth day (D5) mice were sacrificed, their bilateral uterine horns, rushed from the embryonic.Statistics each thrust ratio and the total number of embryos hatched blastocysts/take not hatched blastocysts, was used as the index to evaluate embryos to obtain efficiency;Statistical trophectoderm cell number/inner cell mass number, as a reference index to evaluate the quality of embryo.The results found that groups of Ⅰ, Ⅱ and Ⅲ were no significant difference in the blastocyst number (P>0.05), but there was increasing trend, group Ⅳ was significantly higher than the other three groups (P<0.05).Groups Ⅰ, Ⅱ, Ⅲ and Ⅳ within the cell mass cells number/trophectoderm cell number were 23.18%, 23.55%, 21.72% and 23.28%, there was no significant difference in each group (P>0.05).The results showed that the corresponding set of Ⅳ cage got the time of mouse hatched blastocysts to obtain the highest efficiency, there was no significant differences in embryonic blastocyst quality.

Key words: mouse; hatched blastocysts; superovulation; cage time

中图分类号:

S814.8

翟椿东, 顾美超, 刘迪, 王力红, 齐晓龙, 张嘉楠, 刘云海, 倪和民, 郭勇. 通过优化超排后合笼时间高效获取小鼠孵化囊胚[J]. , 2015, 42(5): 1253-1258.

ZHAI Chun-dong, GU Mei-chao, LIU Di, WANG Li-hong, QI Xiao-long, ZHANG Jia-nan, LIU Yun-hai, NI He-min, GUO Yong. Study on Time Period Model for Efficiently Collecting Hatched Blastocysts Flushed from Murine Uterus[J]. , 2015, 42(5): 1253-1258.

参考文献

[1] Papaioannou V E, Ebert K M.Development of fertilized embryos transferred to oviducts of immature mice[J].J Reprod Fertil, 1986, 76(2):603-608.
[2] Kaufman M H, Barton S C, Surani M A.Normal postimplantation development of mouse parthenogenetic embryos to the forelimb bud stage[J].Nature, 1977, 265(5589):53-55.
[3] Harlow G M, Quinn P.Fetal and placental growth in the mouse after preimplantation development in vitro under oxygen concentrations of 5 and 20%[J].Aust J Biol Sci, 1979, 32(3):363-369.
[4] Wang H, Dey S K.Roadmap to embryo implantation:Clues from mouse models[J].Nature Reviews Genetics, 2006, 7(3):185-199.
[5] 陈琦, 彭洪英, 张莹, 等.胚胎植入研究的回顾和展望[J].科学通报, 2009, 54(18):2771-2782.
[6] Finn C A, Martin L.The control of implantation[J]. Reprod Fertil, 1974, 39:195-206.
[7] Song H, Lim H, Paria B C, et al.Cytosolic phospholipase A2alpha is crucial [correction of A2alpha deficiency is crucial] for 'on-time'embryo implantation that directs subsequent development[J].Development, 2002, 129(12):2879-2889.
[8] Finn C A, Martin L.The control of implantation[J].Reprod Fertil, 1974, 39:195-206.
[9] Paria B C, Huet-hudson Y M, Dey S K.Blastocyst's state of activity determines the "window" of implantation in the receptive mouse uterus[J]. Proc Natl Acad Sci USA, 1993, 90:10159-10162.
[10] Wang H, Tranguch S, Xie H, et al.Variation in commercial rodent diets induces disparate molecular and physiological changes in the mouse uterus[J].Proc Natl Acad Sci USA, 2005, 102(28):9960-9965.
[11] Handyside A H, Hunter S.A rapid procedure forvisualising the inner cell mass and trophectoderm nuclei of mouse blastocysts in situ using polynudeotide-specific fluorochromes[J].J Exp Zool, 1984, 231(3):429-434.
[12] De la Fuente R, King W A.Use of a chemically defined system for the direct comparison of inner cell mass and trophectoderm distribution in murine, pcrcine and bovine embryos[J].Zygote, 1997, 5(4):309-320.
[13] Thouas G A, Korfiatis N A, French A J, et al.Simplified technique for differential staining of inner cell mass and trophectoderm cells of mouse and bovine blastocysts[J].Reprod Biomed Online, 2001, 3(1):25-29.
[14] 卢天罡, 刘云海, 郭勇, 等.一种鉴定小鼠胚胎质量的双重荧光染色方法[J].动物学报, 2008, 54(5):928-932.
[15] 孟令君, 高建民, 刘云海, 等.双重荧光染色法评定小鼠体内外囊胚质量的研究[J].中国畜牧杂志, 2008, 44(3):49-52.
[16] 李瑞岐, 桑润滋.以囊胚双重染色方法检测果糖对胚胎早期发育效果的影响[J]. 中国畜牧杂志, 2010, 7:24-26.
[17] 孙兴参, 谭景和.哺乳动物胚胎计数的几种方法[J].国外畜牧科技, 1992, 23(5):25-26.
[18] 吴兴, 陈峥嵘, 张光健, 等.AO/EB双重染色法检测人骨肉瘤细胞凋亡[J].肿瘤, 2004, 24(1):88-89.
[19] Farahavar A, Shirazi A, Kohram H, et al.Culture of ovine IVM/IVF zygotes in isolated mouse oviduct:Effect of basal medium[J].Avicenna J Med Biotech, 2013, 5(2):133-137.
[20] Hardy K, Handyside A H, Winston R M, et al.The human blastocyst:Cell numver, death and allocation during the preimplantation development in vitro[J].Develoemnt, 1989, 107(3):597-604.
[21] Juan J T, Vanessa G P, Sonia P A, et al.Predictive variable of in vitro fertilization and preimplantation embryo development of ICR mouse 1-cell embryos[J].Fertil Steril, 2002, 74:996-1000.
[22] Vander E J, Amerijckx Y, Van S A, et al.Ultra-rapid freezing of mouse oocytes lowers the cell number in the inner cell mass of 5 day old in vitro cultured blastocysts[J].Hum Reprod, 1998, 13(6):1595-1599.

通过优化超排后合笼时间高效获取小鼠孵化囊胚

Study on Time Period Model for Efficiently Collecting Hatched Blastocysts Flushed from Murine Uterus

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	张笑梦, 焦安惠, 王玉琪, 方南洙, 金庆国. 添加罗格列酮对小鼠卵母细胞氧化还原稳态的影响[J]. 中国畜牧兽医, 2023, 50(5): 1818-1827.
[2]	陆辉, 蔡高峰, 武彩红, 邱树磊, 袁橙, 陈晓兰, 刘芸, 覃凯莉. 茶树菇多糖对小鼠脾脏淋巴细胞免疫调节的影响[J]. 中国畜牧兽医, 2023, 50(5): 1828-1835.
[3]	邵永斌, 郑一闯, 张竞怡, 魏琳颖, 田勇, 孙国结, 刘雨然, 罗燕, 谷新利. 益母草碱对LPS诱导小鼠子宫内膜炎的抗炎作用研究[J]. 中国畜牧兽医, 2023, 50(5): 2149-2155.
[4]	郭雅茹, 李可欣, 王丽, 刘锦钰, 石磊, 任有蛇. 硒对铅致小鼠睾丸间质细胞凋亡的影响[J]. 中国畜牧兽医, 2023, 50(4): 1404-1411.
[5]	史宏昭, 刘洋, 姚睿智, 刘鹏, 李士泽. 冷应激复温对ICR小鼠神经行为学的影响[J]. 中国畜牧兽医, 2023, 50(3): 932-941.
[6]	徐倩倩, 杨万群, 王玉波, 吴菲, 代娟娟, 秦蜜蜜, 庄金秋, 王艳, 沈志强, 李继昌. 熟地黄多糖对环磷酰胺诱导的免疫抑制小鼠的免疫调节作用研究[J]. 中国畜牧兽医, 2023, 50(2): 693-703.
[7]	王巍, 李春洙, 王沙沙, 桑锐, 葛冰洁, 李春亭, 赵欣, 刘馨蔓, 于明弘, 张雪梅. 中药联合益生菌对大肠杆菌致小鼠腹泻保护机制研究[J]. 中国畜牧兽医, 2023, 50(2): 779-788.
[8]	李佳豪, 李超, 臧长江, 孟军, 李宏坤, 贾怡琪, 麦丽克扎提·艾尔肯, 唐纳尔·托留别克. 灌胃和腹腔注射鼠李糖乳杆菌对妊娠期小鼠粪便菌群多样性的影响[J]. 中国畜牧兽医, 2023, 50(1): 151-160.
[9]	于凯, 赵玉芬, 翁钰, 王琴, 郝绍瑜, 于泊洋, 杜晨光, 苏布登格日勒, 李海军. 双氢睾酮对小鼠卵泡颗粒细胞增殖与抗苗勒管激素表达的影响[J]. 中国畜牧兽医, 2022, 49(8): 3036-3043.
[10]	李春亭, 王沙沙, 赵欣, 王巍, 葛冰洁, 张雪梅. 中药联合益生菌对大肠杆菌性小鼠腹泻的保护作用[J]. 中国畜牧兽医, 2022, 49(8): 3226-3234.
[11]	王志亮, 杨喜喜, 铁雅楠, 伏晓玉, 王欣荣. 超排处理对家兔生殖器官形态及激素水平的影响[J]. 中国畜牧兽医, 2022, 49(6): 2250-2258.
[12]	邱树磊, 陈晓兰, 杨海峰, 陈玉库, 胡元亮, 高永旭, 武彩红. 硒化大蒜多糖对小鼠腹腔巨噬细胞功能的影响[J]. 中国畜牧兽医, 2022, 49(3): 1144-1152.
[13]	田微, 方妍雅, 柳海星, 李钟淑. bFGF对3-硝基丙酸处理的小鼠受精卵体外发育能力的影响[J]. 中国畜牧兽医, 2022, 49(2): 608-614.
[14]	张园园, 田志辰, 孙雨佳, 张慧敏, 杨章平, 王聪, 曹柯, 毛永江, 李明勋. 补充牛磺酸对高脂饮食C57BL/6J小鼠糖脂代谢的影响[J]. 中国畜牧兽医, 2022, 49(12): 4604-4613.
[15]	郭子萌, 王福正, 王承志, 杜鹏, 张明月, 李泽垚, 苏晓晨, 吴高峰, 杨建成. 牛磺酸复合添加剂对免疫低下小鼠免疫功能的影响[J]. 中国畜牧兽医, 2022, 49(12): 4635-4645.