囊胚培养液对小鼠胚胎的毒性:体外生殖辅助医疗器械安全性评价
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摘要
背景:囊胚培养液是可以支持胚胎从受精卵发育到囊胚阶段的一种重要培养液,其安全性直接影响囊胚的质量。目的:观察囊胚培养液对小鼠胚胎发育的影响。方法:选用B6D2F1品系小鼠,将雌性小鼠进行超数排卵处理,与同品系雄鼠合笼过夜,次日收集1细胞期受精卵,在M16培养液中培养至4细胞期后,检查胚胎发育情况并转移到受试囊胚培养液中继续培养(实验组),5 d后检查囊胚发育情况并计算囊胚形成率。同时,用含有内毒素的囊胚培养液M16培养小鼠受精卵,作为阳性对照组;用已被证实没有胚胎毒性的囊胚培养液M16培养液培养1细胞期受精卵,作为阴性对照组。结果与结论:阳性对照组囊胚形成率为0,阴性对照组囊胚形成率为87.1%,实验组囊胚形成率为87.3%,实验组囊胚形成率大于80%,说明受试囊胚培养液可以支持小鼠受精卵正常发育到囊胚阶段,对胚胎没有毒性效应。
BACKGROUND: The blastula culture medium can assist the development of zygote from the fertilized egg to the blast blastula. The safety and quality of blastula culture medium directly influences the quality of blastula. OBJECTIVE: To evaluate the effect of blastula culture medium on the development of mouse embryos. METHODS: In this study, B6D2F1 mice were used. The female mice were superovulated and mated with male B6D2F1 mice. One day later, the zygotes were collected and cultured in the M16 medium to 4-cell stage. Then, 4-cell stage embryos were transferred into the tested blastula culture medium(experimental group). After 5 days of culture, the forming rate of blastula was examined. Meanwhile, the M16 medium containing endotoxin was used to culture 1-cell mouse zygote(positive control group). The M16 medium with no embryo toxicity was used to culture 1-cell zygote(negative control group). RESULTS AND CONCLUSION: The formation rate of blastula was 0 in the positive group, 87.1% in the negative control group, and 87.3% in the experimental group. From the results, the tested blastula culture medium could assist the 1-cell zygote growing to the stage of blastula, and the formation rate of blastula was above 80%. The tested blastula culture medium had no toxicity to the mouse embryo.
BACKGROUND: The blastula culture medium can assist the development of zygote from the fertilized egg to the blast blastula. The safety and quality of blastula culture medium directly influences the quality of blastula. OBJECTIVE: To evaluate the effect of blastula culture medium on the development of mouse embryos. METHODS: In this study, B6D2F1 mice were used. The female mice were superovulated and mated with male B6D2F1 mice. One day later, the zygotes were collected and cultured in the M16 medium to 4-cell stage. Then, 4-cell stage embryos were transferred into the tested blastula culture medium(experimental group). After 5 days of culture, the forming rate of blastula was examined. Meanwhile, the M16 medium containing endotoxin was used to culture 1-cell mouse zygote(positive control group). The M16 medium with no embryo toxicity was used to culture 1-cell zygote(negative control group). RESULTS AND CONCLUSION: The formation rate of blastula was 0 in the positive group, 87.1% in the negative control group, and 87.3% in the experimental group. From the results, the tested blastula culture medium could assist the 1-cell zygote growing to the stage of blastula, and the formation rate of blastula was above 80%. The tested blastula culture medium had no toxicity to the mouse embryo.
引文
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[21]邓彬,李予,王文军,等.胚胎培养液氨基酸浓度与体外受精-胚胎移植妊娠结局的关系[J].中山大学学报:医学科学版,2013,24(4):900-905.
[22]胡煜,刘吉,李宝山,等.人类辅助生殖技术中DAY3胚胎质量与囊胚形成相关性分析[J].中国优生与遗传杂志,2012,22(7):114-116.
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[29]舒金辉,张波,冯贵雪.人类囊胚期胚胎培养-移植的研究现状[J].中国比较医学杂志,2009,19(9):69-72.
[30]张宁,王喜良,李侠,等.鼠胚生物学分析在IVF实验室质量控制中的应用[J].中国优生与遗传杂志,2005,15(7):123-124.
[31]胡泊,孙云霞,史敏,等.序贯培养对小白鼠胚胎体外发育的影响[J].新疆医科大学学报,2003,26(4):368-369.
[2]陈贵安.北京大学第三医院开展辅助生育技术近10年回顾[J].北京大学学报:医学版,2002,34(5):623-626.
[3]庄广伦,卢光琇,石一复,等.辅助生育技术进展[J].中国实用妇科与产科杂志,2001,17(1):6-31.
[4]韩堃,郑源强,石艳春.人类辅助生殖技术研究进展[J].中国妇幼健康研究,2008,19(6):606-609.
[5]张婷,王晓民.体外受精与试管婴儿——2010年诺贝尔生理学或医学奖简介[J].首都医科大学学报,2010,30(5):678-682.
[6]彭靖,卢大儒.试管婴儿技术的发展与探讨[J].自然杂志,2010,32(6):338-343,308.
[7]乔杰,马彩虹.中国辅助生殖技术发展现状[J].国际生殖健康/计划生育杂志,2012,31(5):332-333.
[8]赵杰,陈秀娟,范文斌.鼠胚在人类辅助生殖培养环境质量控制的作用研究[J].内蒙古医学院学报,2009,31(1):49-51.
[9]王增艳,何方方,孙正怡,等.冷冻环玻璃化法冷冻小鼠卵母细胞的效果评价[J].生殖医学杂志,2008,17(1):38-42.
[10]朱健生,季钢,张玲,等.囊胚序贯培养在体外受精-胚胎移植中的应用[J].中国优生与遗传杂志,2008,18(8):109-110.
[11]周黎明.囊胚培养及移植在体外受精过程中的临床应用[J].现代实用医学,2008,20(7):553-554.
[12]孙新明,张建平,孙弋.山羊体外受精胚胎序贯培养的研究[J].生物技术,2008,18(4):49-51.
[13]黄志辉,余敏,伍琼芳,等.囊胚培养的相关因素分析[J].江西医药,2013,48(4):305-308.
[14]晁贺,刘英,王树玉,等.序贯培养对比序贯联合子宫内膜细胞共培养的方法对体外胚胎发育的影响[J].中国优生与遗传杂志,2013,23(9):96-98.
[15]丁芳,李志英,周红林,等.小鼠早期胚胎几种不同培养方法的比较[J].重庆医学,2010,39(8):907-909.
[16]郝桂琴,李蓉,耿岚,等.囊胚培养及囊胚移植[J].中国组织工程研究与临床康复,2010,14(31):5801-5804.
[17]李军锋,潘红梅,丁红雷,等.兔原核胚体外序贯培养[J].中国兽医学报,2004,24(3):289-291.
[18]王艳玲,曹云霞.胚胎体外培养的研究进展[J].国外医学:妇幼保健分册,2004,15(4):219-222.
[19]杨健之,刘嘉茵,洪蕾.囊胚培养在体外受精和胚胎移植周期中的应用[J].江苏医药,2000,26(5):340-341,415.
[20]铁新琴,杨如镜.囊胚期与卵裂期胚胎培养与移植比较[J].医药论坛杂志,2006,27(10):3-5.
[21]邓彬,李予,王文军,等.胚胎培养液氨基酸浓度与体外受精-胚胎移植妊娠结局的关系[J].中山大学学报:医学科学版,2013,24(4):900-905.
[22]胡煜,刘吉,李宝山,等.人类辅助生殖技术中DAY3胚胎质量与囊胚形成相关性分析[J].中国优生与遗传杂志,2012,22(7):114-116.
[23]Gardner DK,Reed L,Linck D,et al.Quality control in Human IVF.Sem ReprodM ed.2005;23:319-324.
[24]Gardner DK,Lane M.Embryo Culture Systems in InV itro Fertilization:A Practical Approach.Informa Health Care,NY,2007:221-282.
[25]Weiss TJ,Warnes GM,Gardner DK.Mouse Embryos and Quality Control in Human IVF.Reprod Fertil Dev.1992;4:105-7.
[26]Davidson A,Vermesh M,Lobo RA,et al.Mouse embryo culture as quality control for human in vitro fertilization:the one-cell versus two-cell model.Fert Steril.1988;49:514-521.
[27]Obstetric and gynecologic devices;reclassification and classification of medical devices used for in vitro fertilization and related assisted reproduction procedures--FDA.Final rule.Fed Regist.1998;63(175):48428-48437.
[28]Institute of Laboratory Animal Resources Commission on Life Sciences National Research Council.Guide for the Care and Use of Laboratory Animals.National Academy Press,Washington,1996.
[29]舒金辉,张波,冯贵雪.人类囊胚期胚胎培养-移植的研究现状[J].中国比较医学杂志,2009,19(9):69-72.
[30]张宁,王喜良,李侠,等.鼠胚生物学分析在IVF实验室质量控制中的应用[J].中国优生与遗传杂志,2005,15(7):123-124.
[31]胡泊,孙云霞,史敏,等.序贯培养对小白鼠胚胎体外发育的影响[J].新疆医科大学学报,2003,26(4):368-369.