新西兰兔卵母细胞体外成熟、体外受精及受精卵体外培养的研究
|
摘要
本实验以44只新西兰母兔为实验素材,主要研究兔卵巢表面卵母细胞体外成熟、体外受精以及受精卵的不同体外培养条件,以优化兔卵母细胞体外成熟、体外受精以及受精卵培养的体系,实验结果如下:
1.采用切割法、针刺挤压法收集卵巢表面卵母细胞,检验不同采集方法对收集卵巢卵母细胞数量的影响。结果表明:针刺挤压法的所获得的卵母细胞数(23.63/只)显著高于切割法(16.50/只)(p<0.05)。
2.比较添加不同浓度的FSH、LH对兔卵母细胞体外成熟的影响,结果表明: 1 U /mL FSH+2 U/mL LH的成熟率最高,达到46. 62 %,极显著高于对照组(TCM199+10 %FCS)的成熟率(13. 71 %,p<0.01),显著高于高浓度组(10 U /mL FSH+20 U/mL LH)的成熟率(26.83 %,p<0.05)。
3.比较了不同的成熟培养时间对卵母细胞成熟率的影响,以成熟培养液培养卵母细胞,分别在成熟培养24 h、30 h、36 h、40 h对部分卵母细胞用透明质酸酶处理后观察第一极体排出率,其余部分加精子观察受精率。培养24 h、30 h、36 h、40 h的成熟率分别为21.92 %、46.62 %、52.77 %、30.71 %,这就表明:成熟时间为30、36 h的成熟率显著高于成熟时间为24、40 h的成熟率(p<0.05)。
4.以TCM199+FCS+FSH+LH+E2+丙酮酸钠为基础成熟培养液,分别添加50 ng/mL及100 ng/mL的EGF、10 ng/mL及20 ng/mL的SCF、10 ng/mL及20 ng/mL的LIF,然后比较了EGF、SCF及LIF对卵母细胞成熟率的影响。结果表明:50 ng/mL及100 ng/mL的EGF能显著地提高了卵母细胞成熟率(基础成熟培养液、添加50 ng/mL及100 ng/mL的EGF组的成熟率分别为49.04 %、61.39 %和65.79 %)。SCF对卵母细胞成熟率无明显影响(成熟率分别为54.17 %、50.00 %和45.83 %),而LIF对卵母细胞成熟有明显的抑制作用(成熟率分别为54.19 %、12.50 %和4.17 %)。
5.比较了上游法、直接洗涤法和Percoll密度梯度离心法三种精子处理方法对兔精子回收率及卵裂率的影响,三种方法的回收率分别为7.8 %、93.9 %、86.9 %,受精后的卵裂率分别为0 %、39.05 %、54.29 %,结果表明:Percoll密度梯度离心法是一种较理想的精子处理方法。
6.比较了常规体外受精与显微注射卵裂率,结果显示,常规体外受精卵裂率(63.97 %)极显著高于显微注射卵裂率(25.65 %)(p<0.01)。
7.比较了TCM199+10% FCS和DMEM+10 %FCS对胚胎(取自常规体外受精的2-细胞)发育的影响,结果表明两者之间无显著差异(p>0.05)。
8.以TCM199+10 %FCS为基础的受精卵培养液,分别添加一定浓度的维生素C(100μmol/L)及β-巯基乙醇(50μmol/L),然后比较了它们对胚胎(取自常规体外受精的2-细胞)发育的影响,表明结果,100μmol/L的维生素C及50μmol/Lβ-巯基乙醇对胚胎发育无显著影响(p>0.05)。
In this paper , we took 44 female New Zealand Rabbits as object, mainly researched in vitro maturation,fertilization of oocyte on ovarian surface and in vitro culture of zygote,our purposes were optimizing the system of in vitro maturation,fertilization of rabbit oocyte and in vitro culture of zygote,results were as follows.
1. Cumulus and oocyte complex (COCs) on the surface of ovaries were gotten by the method of cutting with Blade or stab-extruding with needle,the number of COCs gotten by the method of stab-extruding with needle(23.63 per rabbit )were significant larger than by the method of cutting (16.50 per rabbit )(P <0.05).
2. Supplemented with diffirent concentration of FSH+LH in the maturation medium,then COCs were IVM under the condition of 38.5°C,5 % CO2 and saturated humidity,the time of IVM was 30 h,the results showed that the rate of maturation by supplementing 1 U/mL FSH+2 U/mL LH was significent higher than 10 U/mL FSH+20 U/mL LH,and more significent higher than contrast group(TCM199+10 %FCS).The rate of maturation were respectively 46. 62 %,26.83 %,13. 71 %.
3. This experiment compared the rate of maturation of oocytes by the time of 24 h,30 h,36 h or 40 h,the results indicated that culture time of 30 h or 36 h were better than 24 h or 40 h(P <0.05).The rate of maturation were 21.92 %,46.62 %,52.77 %, 30.71 % respectively.
4. Basic maturation medium was TCM199+FCS+FSH+LH+E2+pyruvic sodium,added epidermal growth factor (EGF)50 ng/mL,EGF 100ng/mL,stem cell factor (SCF)10 ng/mL,SCF 20 ng/mL,leukemia inhibitory factor(LIF)10 ng/mL,LIF 20 ng/mL,then counted rate of first polar body emission by the time of 30 h .Results manifested the first polar body emission rate in group of Adding 50 ng / mL and 100 ng / mL EGF were significantly higher(P <0.05),first polar body emission rate of contrast group,50 ng / mL group and 100 ng / mL was respectively 49.04 %,61.39 % and 65.79 %,SCF could not enhance the first polar body emission rate,first polar body emission rate was respectively 54.17 %,50.00 % and 45.83 %,LIF obviously restrained the maturation of oocyte,first polar body emission rate was respectively 54.19 %,12.50 % and 4.17 %.
5. Test compared three methods of sperm handlling on sperm recovery rate and cleavage rate , they were swimming-up , direct washing and density gradient centrfugation,results showed that the method of density gradient centrfugation was a ideal method(sperm recovery rate of the three ways were 7.8 %,93.9 % and 86.9 %,cleavage rate after the fertilization were 0 %,39.05 % and 54.29 %).
6. Cleavage rate of in vitro fertilization and cleavage rate of intracytoplasmic sperm injection was compared,result was that the cleavage rate of former(63.97 %)was more significant higher than the later(25.65 %)(P <0. 01).
7. The experiment compared embryo developmental potential capacity between TCM199+10 %FCS group and DMEM+10 %FCS group,the result indicated that they have no significant difference(P >0. 05).
8. Supplemented with vitmin C(100μmol/L)and mercaptoethanol (50μmol/L)in the zygote culture medium,zygote of 2-cell (from in vitro fertilization)were cultured under the condition of 38.5℃,5% CO2 and saturated humidity,then statisticed the rate of 4-cell and of 8-cell or later,the results were that this three groups have no significat difference,Whether the rate of 4-cell zygote or rate of 8-cell zygote or later.
1.采用切割法、针刺挤压法收集卵巢表面卵母细胞,检验不同采集方法对收集卵巢卵母细胞数量的影响。结果表明:针刺挤压法的所获得的卵母细胞数(23.63/只)显著高于切割法(16.50/只)(p<0.05)。
2.比较添加不同浓度的FSH、LH对兔卵母细胞体外成熟的影响,结果表明: 1 U /mL FSH+2 U/mL LH的成熟率最高,达到46. 62 %,极显著高于对照组(TCM199+10 %FCS)的成熟率(13. 71 %,p<0.01),显著高于高浓度组(10 U /mL FSH+20 U/mL LH)的成熟率(26.83 %,p<0.05)。
3.比较了不同的成熟培养时间对卵母细胞成熟率的影响,以成熟培养液培养卵母细胞,分别在成熟培养24 h、30 h、36 h、40 h对部分卵母细胞用透明质酸酶处理后观察第一极体排出率,其余部分加精子观察受精率。培养24 h、30 h、36 h、40 h的成熟率分别为21.92 %、46.62 %、52.77 %、30.71 %,这就表明:成熟时间为30、36 h的成熟率显著高于成熟时间为24、40 h的成熟率(p<0.05)。
4.以TCM199+FCS+FSH+LH+E2+丙酮酸钠为基础成熟培养液,分别添加50 ng/mL及100 ng/mL的EGF、10 ng/mL及20 ng/mL的SCF、10 ng/mL及20 ng/mL的LIF,然后比较了EGF、SCF及LIF对卵母细胞成熟率的影响。结果表明:50 ng/mL及100 ng/mL的EGF能显著地提高了卵母细胞成熟率(基础成熟培养液、添加50 ng/mL及100 ng/mL的EGF组的成熟率分别为49.04 %、61.39 %和65.79 %)。SCF对卵母细胞成熟率无明显影响(成熟率分别为54.17 %、50.00 %和45.83 %),而LIF对卵母细胞成熟有明显的抑制作用(成熟率分别为54.19 %、12.50 %和4.17 %)。
5.比较了上游法、直接洗涤法和Percoll密度梯度离心法三种精子处理方法对兔精子回收率及卵裂率的影响,三种方法的回收率分别为7.8 %、93.9 %、86.9 %,受精后的卵裂率分别为0 %、39.05 %、54.29 %,结果表明:Percoll密度梯度离心法是一种较理想的精子处理方法。
6.比较了常规体外受精与显微注射卵裂率,结果显示,常规体外受精卵裂率(63.97 %)极显著高于显微注射卵裂率(25.65 %)(p<0.01)。
7.比较了TCM199+10% FCS和DMEM+10 %FCS对胚胎(取自常规体外受精的2-细胞)发育的影响,结果表明两者之间无显著差异(p>0.05)。
8.以TCM199+10 %FCS为基础的受精卵培养液,分别添加一定浓度的维生素C(100μmol/L)及β-巯基乙醇(50μmol/L),然后比较了它们对胚胎(取自常规体外受精的2-细胞)发育的影响,表明结果,100μmol/L的维生素C及50μmol/Lβ-巯基乙醇对胚胎发育无显著影响(p>0.05)。
In this paper , we took 44 female New Zealand Rabbits as object, mainly researched in vitro maturation,fertilization of oocyte on ovarian surface and in vitro culture of zygote,our purposes were optimizing the system of in vitro maturation,fertilization of rabbit oocyte and in vitro culture of zygote,results were as follows.
1. Cumulus and oocyte complex (COCs) on the surface of ovaries were gotten by the method of cutting with Blade or stab-extruding with needle,the number of COCs gotten by the method of stab-extruding with needle(23.63 per rabbit )were significant larger than by the method of cutting (16.50 per rabbit )(P <0.05).
2. Supplemented with diffirent concentration of FSH+LH in the maturation medium,then COCs were IVM under the condition of 38.5°C,5 % CO2 and saturated humidity,the time of IVM was 30 h,the results showed that the rate of maturation by supplementing 1 U/mL FSH+2 U/mL LH was significent higher than 10 U/mL FSH+20 U/mL LH,and more significent higher than contrast group(TCM199+10 %FCS).The rate of maturation were respectively 46. 62 %,26.83 %,13. 71 %.
3. This experiment compared the rate of maturation of oocytes by the time of 24 h,30 h,36 h or 40 h,the results indicated that culture time of 30 h or 36 h were better than 24 h or 40 h(P <0.05).The rate of maturation were 21.92 %,46.62 %,52.77 %, 30.71 % respectively.
4. Basic maturation medium was TCM199+FCS+FSH+LH+E2+pyruvic sodium,added epidermal growth factor (EGF)50 ng/mL,EGF 100ng/mL,stem cell factor (SCF)10 ng/mL,SCF 20 ng/mL,leukemia inhibitory factor(LIF)10 ng/mL,LIF 20 ng/mL,then counted rate of first polar body emission by the time of 30 h .Results manifested the first polar body emission rate in group of Adding 50 ng / mL and 100 ng / mL EGF were significantly higher(P <0.05),first polar body emission rate of contrast group,50 ng / mL group and 100 ng / mL was respectively 49.04 %,61.39 % and 65.79 %,SCF could not enhance the first polar body emission rate,first polar body emission rate was respectively 54.17 %,50.00 % and 45.83 %,LIF obviously restrained the maturation of oocyte,first polar body emission rate was respectively 54.19 %,12.50 % and 4.17 %.
5. Test compared three methods of sperm handlling on sperm recovery rate and cleavage rate , they were swimming-up , direct washing and density gradient centrfugation,results showed that the method of density gradient centrfugation was a ideal method(sperm recovery rate of the three ways were 7.8 %,93.9 % and 86.9 %,cleavage rate after the fertilization were 0 %,39.05 % and 54.29 %).
6. Cleavage rate of in vitro fertilization and cleavage rate of intracytoplasmic sperm injection was compared,result was that the cleavage rate of former(63.97 %)was more significant higher than the later(25.65 %)(P <0. 01).
7. The experiment compared embryo developmental potential capacity between TCM199+10 %FCS group and DMEM+10 %FCS group,the result indicated that they have no significant difference(P >0. 05).
8. Supplemented with vitmin C(100μmol/L)and mercaptoethanol (50μmol/L)in the zygote culture medium,zygote of 2-cell (from in vitro fertilization)were cultured under the condition of 38.5℃,5% CO2 and saturated humidity,then statisticed the rate of 4-cell and of 8-cell or later,the results were that this three groups have no significat difference,Whether the rate of 4-cell zygote or rate of 8-cell zygote or later.
引文
[1] Pincus G, Enzmann E V. The comparative behavior of mammalian eggs in vivo and in vitro : the activation of ovarian [J]. J Exp Med. 1935, 62(5): 665-675.
[2] Pincus G. Ovum culture[J]. Science. 1939, 89(2318): 509.
[3] Al-Hasani S, Trotnow S, Sadtler C, et al. In vitro fertilization and embryo transfer of pre-ovulatory rabbit oocytes[J]. Eur J Obstet Gynecol Reprod Biol. 1986, 21(3): 187-195.
[4] S C E, L L M, H E W. Acquisition of developmental competence during maturation in vitro [J]. Theriogenology. 1986(25): 150.
[5] Dostal J, Pavlok A. Isolation and characterization of maturation inhibiting compound in bovine follicular fluid[J]. Reprod Nutr Dev. 1996, 36(6): 681-690.
[6] Armstrong D T, Irvine B J, Earl C R, et al. Gonadotropin stimulation regimens for follicular aspiration and in vitro embryo production from calf oocytes[J]. Theriogenology. 1994, 42(7): 1227-1236.
[7]麻柱,桑润滋.牛活体采卵技术研究进展[J].中国畜牧杂志. 2006, 42(03): 64-66.
[8]唐修君,陈丽,邹海军,等.家兔卵母细胞体外受精研究[J].中国畜牧杂志. 2007, 43(15): 15-17.
[9] Mayes M A, Sirard M A. The influence of cumulus-oocyte complex morphology and meiotic inhibitors on the kinetics of nuclear maturation in cattle[J]. Theriogenology. 2001, 55(4): 911-922.
[10] Otoi T, Yamamoto K, Koyama N, et al. Cryopreservation of mature bovine oocytes by vitrification in straws[J]. Cryobiology. 1998, 37(1): 77-85.
[11]翟中和,王喜忠,丁明孝主编.细胞生物学[M].北京:高等教育出版社, 2001.
[12]韩树标,王爱萍,张力,等. MPF和MAPK在卵母细胞成熟过程中的作用[J].吉林畜牧兽医. 2007, 28(5): 14-16.
[13] T K, K N, Sugiura K E A. Analysis of the roles of cyclin B1 and cyclin B2 in porcine oocyte maturation by inhibiting synthesis with antisense RNA injection[J]. Biol Reprod. 2004, 70(1):154-159.
[14] S B, B M. Cytoskeleton and cell cycle control during meiotic maturation of the mouse oocyte : integrating time and space[J]. Reproduction. 2005, 130(6): 801-811.
[15] Kim J S, Cho Y S, Song B S, et al. Exogenous dibutyryl cAMP affects meiotic maturation via protein kinase A activation; it stimulates further embryonic development including blastocyst quality in pigs[J]. Theriogenology. 2008, 69(3): 290-301.
[16] Rg M, Jp N, Pb G. Protein kinase C(PKC) role in bovine oocyte maturation and early embryo development[J]. Anim Reprod Sci. 2008, 107(1-2): 20-29.
[17] Yh Z, Lp Z, Li F E A. C-erbB2 and c-myb induce oocyte maturation via activation of mitogen-activated protein kinase and maturation promoting factor[J]. Acta Physiol Sin. 2008, 50(1): 97-104.
[18] Liang C G, Huo L J, Zhong Z S, et al. Cyclic adenosine 3',5'-monophosphate-dependent activation of mitogen-activated protein kinase in cumulus cells is essential for germinal vesicle breakdown of porcine cumulus-enclosed oocytes[J]. Endocrinology. 2005, 146(10): 4437-4444.
[19] Hansen D V, Pomerening J R, Summers M K, et al. Emi2 at the crossroads: where CSF meets MPF[J]. Cell Cycle. 2007, 6(6): 732-738.
[20] M O, D I, Kanemori Y E A. Erp1 /Emi2 is essential for the meiosis I to meiosis II transition in Xenopus oocytes[J]. Dev Biol. 2007, 303(1): 157-164.
[21] Ito J, Kato M, Hochi S, et al. Effect of enucleation on inactivation of cytostatic factor activity in matured rat oocytes[J]. Cloning Stem Cells. 2007, 9(2): 257-266.
[22] Gotze M, Kauffold P, Schuffenhauer A, et al. The inhibition of meiosis of bovine oocytes using biologic of synthetic inhibitors[J]. Arch Exp Veterinarmed. 1990, 44(1): 19-27.
[23]陈大元.受精生物学-受精机制与生殖工程[M].北京:科学出版社, 2000.
[24]王红铃.不同添加物对牛卵母细胞体外成熟、体外受精及受精卵体外培养的影响[D].南京农业大学, 2002.
[25] Gordon I. Laboratory production of cattle embryos[M]. UK: Commonwealth Agricultural Bureau International, Wallingford, Oxon, 1994.
[26] Jc J, Th C, Jk T, et al. Cytoskeletal patterns, in vitro maturation and parthenogenetic development of rabbit GV oocytes[J].Asian-Australasian Journal of Animal Sciences. 2002, 15(12): 1695-1701.
[27] Tao Y, Cao C, Zhang M, et al. Effects of cumulus cells on rabbit oocyte in vitro maturation.[J]. J Anim Physiol Anim Nutr (Berl). 2008, 92(4): 438-447.
[28] Wtk C, Rm M, C P. In vitro fertilization of pig and sheep oocytes matured in vivo[J]. Theriogenology. 1986, 25: 146.
[29] Lu K H, Gordon I, Gallagher M, et al. Pregnancy established in cattle by transfer of embryos derived from in vitro fertilisation of oocytes matured in vitro[J]. Vet Rec. 1987, 121(11): 259-260.
[30]郑瑞珍,高晓虹,王育哲,等.兔卵母细胞体外成熟和体外受精的研究[J].动物学报. 1992, 38(01): 66-72.
[31]赵晓娥,李向臣,高立功,等.山羊卵母细胞体外成熟研究[J].中国农学通报. 2005, 21(09): 1-4.
[32]索永善,李跃民,华云芳,等. PH值对猪卵母细胞体外成熟的影响[J].畜牧兽医杂志. 2005, 24(02): 4-5.
[33] Eppig J J. Maintance of meiotic arrest and the induction of oocyte maturation in mouse oocyte granulosa cell complexes developed in vitro from preantal follieles [J]. biol Reprod. 1991, 45: 824-830.
[34] Lu K H, Gordon I, Gallagher M, et al. Pregnancy established in cattle by transfer of embryos derived from in vitro fertilization of oocytes matured in vitro[J]. Vet Rec. 1987, 121(11): 259-260.
[35] Bd B, Ta R, T P. Development of in vitro matured in vitro fertilized bovine embryos into morulae and blastocysts in defined cu1ture media[J]. Theriogenology. 1992, 37(1): 127-146.
[36] Dag I, Ab S, Ec L. Deactivation of the serum prior to medium supplementation:a necessit or a myth[J]. Human Reprod. 1992, 7(supple2): 159.
[37]丁爱军,周艳华,贾银海,等.生殖激素对奶水牛卵母细胞成熟及发育潜能的影响[J].中国奶牛. 2008(11): 18-21.
[38]檀利长,田宁宁,霍道坦,等.牛卵泡液对牛卵母细胞体外成熟及受精胚发育力的影响[J].黑龙江动物繁殖. 2008, 16(05): 3-5.
[39] Ikeda S, Kitagawa M, Imai H, et al. The roles of vitamin A for cytoplasmic maturation of bovine oocytes[J]. J Reprod Dev. 2005, 51(1): 23-35.
[40] Dalvit G, Llanes S P, Descalzo A, et al. Effect of alpha-tocopherol and ascorbic acid on bovine oocyte in vitro maturation[J]. Reprod Domest Anim. 2005, 40(2): 93-97.
[41] Yanagimachi R. Gamete manipulation for development: new methods for conception[J]. Reprod Fertil Dev. 2001, 13(1): 3-14.
[42]冯伯森,王秋雨,胡玉兴编著.动物细胞工程原理与实践[M].北京:科学出版社, 2000.
[43]张红卫主编.发育生物学[M].北京:高等教育出版社, 2001.
[44]郭志勤主编.家畜胚胎工程[M].北京:中国科学技术出版社, 1998.
[45]秦鹏春,石惠芝,郭立民.试管牛胚胎的实验室生产过程[J]. 1997.
[46] Totey S M, Pawshe C H, Singh G P. Effects of bull and heparin and sperm concentrations on in vitro fertilization of buffalo (Bubalus bubalis ) oocytes matured in vitro[J]. Theriogenology. 1993, 39(4): 887-898.
[47] Parrish J J, Susko-Parrish J, Winer M A, et al. Capacitation of bovine sperm by heparin[J]. Biol Reprod. 1988, 38(5): 1171-1180.
[48]谭世俭,卢克焕.受精用液种类对牛卵母细胞体外受精的影响[J].中国兽医学报. 1994, 14(02): 140-145.
[49]兰宗保,许惠艳,保若钢,等.精子浓度和授精前孵育时间、精卵共孵育时间对猪卵母细胞体外受精效果的影响[J].广西畜牧兽医.2008,24(3):139-142.
[50]刘健,刘建民,张嘉保.哺乳动物体外受精若干问题[J].中国兽医学报. 1992, 12(03): 307-314.
[51]李子义,周琪,邹啸环,等.家兔显微受精影响因素及显微受精胚胎超微结构[J].中国兽医学报. 1997,(06):593-599.
[52] Kimura Y, Yanagimachi R. Mouse oocytes injected with testicular spermatozoa or round spermatids can develop into normal offspring[J]. Development. 1995, 121(8): 2397-2405.
[53] Wei H, Fukui Y. Fertilizability of ovine, bovine or minke whale (Balaenoptera acutorostrata) spermatozoa intracytoplasmically injected into bovine oocytes[J]. Zygote. 2000, 8(3): 267-274.
[54] Palermo G, Joris H, Devroey P, et al. Pregnancies after intracytoplasmic injection of single spermatozoon into an oocyte[J]. Lancet. 1992, 340(8810): 17-18.
[55] Butler E A, Masson G M. Development of a successful ICSI programme without the use of PVP[J]. Hum Reprod. 1997, 12(5): 1115-1116.
[56] Balaban B, Lundin K, Morrell J M, et al. An alternative to PVP for slowing sperm prior to ICSI[J]. Hum Reprod. 2003, 18(9): 1887-1889.
[57] Goto K. Bovine microfertilization and embryo transfer[J]. Mol Reprod Dev. 1993, 36(2): 288-290.
[58]曾勇,胡晓东,刘洪君,等.卵母细胞浆内单精子注射后电激活的临床应用[J].中华妇产科杂志. 2004, 39(2): 120.
[59]鹿群,陈子江,高选,等.钙离子载体A23187联合嘌呤霉素激活行卵母细胞质内单精子注射后未受精卵母细胞的研究[J].中华妇产科杂志. 2006, 41(3): 182-185.
[60]张晓慧,曹云霞,章志国,等.两种人工激活剂对卵胞浆内单精子注射后未受精卵母细胞激活效果的比较[J].中国妇产科临床杂志. 2009, 10(3): 204-206.
[61] G B, Bracket. Analysis of factors involved in the in vitro Production of bovine embryos[J]. Theriogenology. 1993,39(1): 43-64.
[62] Telford N A, Watson A J, Schultz G A. Transition from maternal to embryonic control in early mammalian development: a comparison of several species[J]. Mol Reprod Dev. 1990, 26(1): 90-100.
[63] Reed M. L编郭年藩摘译.猪胚胎的体外培养[J].国外畜牧科技. 1992, 19(4): 16-18.
[64]曹明聚.绵羊胚胎体外培养条件的研究进展[J].国外畜牧科技. 1996, 23(05): 27-30.
[65]王红玲.不同添加物对牛卵母细胞体外成熟、体外受精、及受精卵体外培养的影响[D].江苏南京:南京农业大学, 2005.
[66] Neira J A, Tainturier D, Pen M A, et al. Effect of the association of IGF-I, IGF-II, bFGF, TGF-b1, GM-CSF,and LIF on the development of bovine embryos produced in vitro[J]. Theriogenology. 2010(73): 595-604.
[67]贾秀芬,周燕华,刘慧雯,等.三种抗氧化剂对小鼠体外受精胚胎发育的影响[J].中国组织化学与细胞化学杂志. 2009, 15(04): 70-73.
[68]张永华.抗氧化剂对小鼠1一细胞期胚胎体外发育的影响[D].吉林:延边大学, 2006.
[69]唐军旺.维生素C和E及血清种类对水牛体外受精早期胚胎发育与凋亡影响的研究[D].广西南宁:广西大学, 2008.
[70]王敏康,刘冀珑,李光鹏,等. M16添加硫磺酸和EDTA支持昆明白小鼠体外受精并发育至囊胚[J].遗传. 2000, 22(05): 301-302.
[71]卢晟盛,魏凤,房慧伶,等.乙二胺四乙酸钠对猪卵母细胞的体外成熟和孤雌激活后早期发育的影响[J].安徽农业大学学报. 2008, 35(01): 9-16.
[72] Lu J H, Wang J Z, Wang H L, et al. Damaging effect of cumulus denudation on rabbit oocytes[J]. Fertil Steril. 2010, 93(5): 1567-1573.
[73]虹卢,杨蓉生,马志杰,等.牛卵母细胞体外成熟的研究进展[J].青海畜牧兽医杂志. 2006, 36(5): 44-45.
[74]戴荣亮,周俊波,王杏龙.山羊卵母细胞体外成熟的研究[J].中国草食动物. 2006, 26(1): 12-14.
[75] Al V S A E. FSH and growth factors affect the growth and endocrine function in vitro of granulose cells of bovine preantral follicles[J]. Theriogenology. 1996(45): 817-832.
[76]刘红林,范必勤.哺乳动物卵母细胞的体外成熟[J].畜牧与兽医. 1996, 28(4): 180-183.
[77]张德福,王建荣.兔卵母细胞体外培养系统的建立[J].上海农业学报. 1994, 10(03): 83-89.
[78]张德福,王济容.家兔卵巢卵母细胞体外成熟和体外受精的初步试验[J].上海农学院学报. 1993, 11(1): 12-16.
[79] L L P, J I. Enhancement of cumulus expansion and unclear maturation during bovine occyte maturation on in vitro by the addition of epidermal growth factor and insulin like growth factor[J]. Fertil, J Rep Rod. 1994, 101(3): 697-701.
[80] Wangw, K N. Syenergetic efficets of EGF factor and gonadotropins on the cytoplasmicmaturation of pig ooctyes in a serum-free medium[J]. Zygot. 1995(3): 345-350.
[81] Kw P. Exposure of bovine oocyte to EGF duringmaturation allows them to develop to blastcysts in a chemically - defined medium[J]. Theriogenology. 1997(48): 1127-1135.
[82]张涌,刘泽隆,李裕.山羊卵泡卵母细胞体外成熟的研究[J].西北农业大学学报. 1999,27(1): 14-18.
[83]冯贵雪,卞桂华,王晓丽,等.表皮生长因子对水牛卵母细胞体外培养核质成熟的影响[J].中国畜牧兽医. 2007, 34(1): 75-78.
[84]贺奋义,桑国俊,余四九,等.牛卵母细胞体外成熟技术研究[J].畜牧兽医杂志. 2008, 27(06): 16-18.
[85]进鲁,刘凤军,王勇胜,等. EGF和IGF - 1对山羊卵母细胞体外成熟的影响[J].黑龙江畜牧兽医. 2005(3): 7-9.
[86]马红,郭镇华,李忠秋,等.表皮生长因子在绵羊卵母细胞体外成熟过程中的作用[J].黑龙江畜牧兽医. 2007(7): 51-52.
[87] Lorenzo P L, Illera M J, Illera J C, et al. Enhancement of cumulus expansion and nuclear maturation during bovine oocyte maturation in vitro by the addition of epidermal growth factor and insulin-like growth factor I[J]. J Reprod Fertil. 1994, 101(3): 697-701.
[88] Grupen C G, Nagashima H, Nottle M B. Role of epidermal growth factor and insulin-like growth factor-I on porcine oocyte maturation and embryonic development in vitro[J]. Reprod Fertil Dev. 1997, 9(6): 571-575.
[89] Smitz J, Cortvrindt R, Hu Y. Epidermal growth factor combined with recombinant human chorionic gonadotrophin improves meiotic progression in mouse follicle-enclosed oocyte culture[J]. Hum Reprod. 1998, 13(3): 664-669.
[90] Lorenzo P L, Liu I K, Carneiro G F, et al. Equine oocyte maturation with epidermal growth factor[J]. Equine Vet J. 2002, 34(4): 378-382.
[91] Purohit G N, Brady M S, Sharma S S. Influence of epidermal growth factor and insulin-like growth factor 1 on nuclear maturation and fertilization of buffalo cumulus oocyte complexes in serum free media and their subsequent development in vitro[J]. Anim Reprod Sci. 2005, 87(3-4): 229-239.
[92] Lorenzo P L, Rebollar P G, Illera M J, et al. Stimulatory effect of insulin-like growth factor I and epidermal growth factor on the maturation of rabbit oocytes in vitro[J]. J Reprod Fertil. 1996, 107(1): 109-117.
[93] Gomez E, de Los S M, Ruiz A, et al. Effects of epidermal growth factor in the final stages of nuclear and cytoplasmic oocyte maturation in humans[J]. Hum Reprod. 1993, 8(5): 691-694.
[94]王炼炼,彦丘,幸贵邦.表皮生长因子促进小鼠未成熟卵母细胞体外成熟的实验研究[J].第三军医大学学报. , 29(22): 2179-2181.
[95]胥焘,窦忠英,陈静波. EGF和GSH对牛卵母细胞体外受精的影响[J].中国农业通报. 2005, 21(6): 55-57.
[96]赵振华,钱红娟,周敏敏,等.细胞因子与激素对山羊卵母细胞体外成熟及卵裂的影响[J].畜牧与兽医. 2008, 40(09): 38-40.
[97] Rodriguez A, De Frutos C, Diez C, et al. Effects of human versus mouse leukemia inhibitory factor on the in vitro development of bovine embryos[J]. Theriogenology. 2007, 67(5): 1092-1095.
[98]周敏敏,丁海雷,钱红娟,等. LIF对小鼠卵母细胞体外成熟和体外受精效果的影响[J].上海畜牧兽医通讯. 2008(03): 38-39.
[99]纪红,刘慧雯,秦逸人,等.显微操作针制作相关问题的探讨[J].生殖医学杂志. 2007, 16(6): 419-422.
[100] Nagy编孙青原陈大元主译.小鼠胚胎操作实验手册[Z].化学工业出版社, 2006.
[101]李嫒.人类辅助生殖实验技术[M].北京:科学出版社, 2008.
[102]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(04): 49-51.
[103]范必勤,熊慧卿,刘铁铮,等.家兔体外受精研究[J].江苏农业学报. 1987, 3(01): 11-17.
[104]任克良,罗惠娣,毛杨毅,等.家兔体外受精技术研究[J].华北农学报. 2000, 15(01): 135-138.
[105]蒋涛,高庆华,何良军,等.兔体外受精的研究[J].中国草食动物. 2005, 25(06): 3-6.
[106] Brackett B G, Mills J A, Jr Jeitles G G. In vitro fertilization of rabbit ova recovered from ovarian follicles[J]. Fertil Steril. 1972, 23(12): 898-909.
[107] Fukuda A, Roudebush W E, Thatcher S S. Platelet activating factor enhances the acrosome reaction, fertilization in vitro by subzonal sperm injection and resulting embryonic development in the rabbit[J]. Hum Reprod. 1994, 9(1): 94-99.
[108]周曾娣,黎曼侬,陈云鹤,等.超声断尾精子头显微受精-输卵管内移植研究的初步报告[J].中国优生与遗传杂志. 2003, 11(01): 72-73.
[109]陈浩杰,刘丽均,张春燕,等.实验兔胞内单精子注射技术[J].中国实验动物学报. 2008, 16(04): 250-253.
[110]唐修君.家兔超数排卵和体外受精技术的研究[D].江苏,扬州:扬州大学, 2007.
[111]李善刚,陈学进,李宏滨,等.不同激活条件和培养基对兔卵母细胞孤雌激活和胚胎发育的影响[J].畜牧兽医学报. 2008, 39(12): 1665-1670.
[112] Wang X, Falcone T, Attaran M, et al. Vitamin C and vitamin E supplementation reduce oxidative stress-induced embryo toxicity and improve the blastocyst development rate[J]. Fertil Steril. 2002, 78(6): 1272-1277.
[113]贾秀芬,郭铁云,周燕华,等.抗氧化剂对小鼠体外受精胚胎发育的影响[J].解剖科学进展. 2009, 15(1): 70-73.
[114]尚江华,张秀芳,黄右军.添加β-巯基乙醇对水牛胚胎体外发育的影响[J].草食家畜. 2001(03): 28-31.
[115] Park E S, Hwang W S, Kang S K, et al. Improved embryo development with decreased apoptosis in blastomeres after the treatment of cloned bovine embryos with beta-mercaptoethanol and hemoglobin[J]. Mol Reprod Dev. 2004, 67(2): 200-206.
[116] Goncalves F S, Barretto L S, Arruda R P, et al. Effect of antioxidants during bovine in vitro fertilization procedures on spermatozoa and embryo development [J]. Reprod Domest Anim. 2010, 45(1): 129-135.
[2] Pincus G. Ovum culture[J]. Science. 1939, 89(2318): 509.
[3] Al-Hasani S, Trotnow S, Sadtler C, et al. In vitro fertilization and embryo transfer of pre-ovulatory rabbit oocytes[J]. Eur J Obstet Gynecol Reprod Biol. 1986, 21(3): 187-195.
[4] S C E, L L M, H E W. Acquisition of developmental competence during maturation in vitro [J]. Theriogenology. 1986(25): 150.
[5] Dostal J, Pavlok A. Isolation and characterization of maturation inhibiting compound in bovine follicular fluid[J]. Reprod Nutr Dev. 1996, 36(6): 681-690.
[6] Armstrong D T, Irvine B J, Earl C R, et al. Gonadotropin stimulation regimens for follicular aspiration and in vitro embryo production from calf oocytes[J]. Theriogenology. 1994, 42(7): 1227-1236.
[7]麻柱,桑润滋.牛活体采卵技术研究进展[J].中国畜牧杂志. 2006, 42(03): 64-66.
[8]唐修君,陈丽,邹海军,等.家兔卵母细胞体外受精研究[J].中国畜牧杂志. 2007, 43(15): 15-17.
[9] Mayes M A, Sirard M A. The influence of cumulus-oocyte complex morphology and meiotic inhibitors on the kinetics of nuclear maturation in cattle[J]. Theriogenology. 2001, 55(4): 911-922.
[10] Otoi T, Yamamoto K, Koyama N, et al. Cryopreservation of mature bovine oocytes by vitrification in straws[J]. Cryobiology. 1998, 37(1): 77-85.
[11]翟中和,王喜忠,丁明孝主编.细胞生物学[M].北京:高等教育出版社, 2001.
[12]韩树标,王爱萍,张力,等. MPF和MAPK在卵母细胞成熟过程中的作用[J].吉林畜牧兽医. 2007, 28(5): 14-16.
[13] T K, K N, Sugiura K E A. Analysis of the roles of cyclin B1 and cyclin B2 in porcine oocyte maturation by inhibiting synthesis with antisense RNA injection[J]. Biol Reprod. 2004, 70(1):154-159.
[14] S B, B M. Cytoskeleton and cell cycle control during meiotic maturation of the mouse oocyte : integrating time and space[J]. Reproduction. 2005, 130(6): 801-811.
[15] Kim J S, Cho Y S, Song B S, et al. Exogenous dibutyryl cAMP affects meiotic maturation via protein kinase A activation; it stimulates further embryonic development including blastocyst quality in pigs[J]. Theriogenology. 2008, 69(3): 290-301.
[16] Rg M, Jp N, Pb G. Protein kinase C(PKC) role in bovine oocyte maturation and early embryo development[J]. Anim Reprod Sci. 2008, 107(1-2): 20-29.
[17] Yh Z, Lp Z, Li F E A. C-erbB2 and c-myb induce oocyte maturation via activation of mitogen-activated protein kinase and maturation promoting factor[J]. Acta Physiol Sin. 2008, 50(1): 97-104.
[18] Liang C G, Huo L J, Zhong Z S, et al. Cyclic adenosine 3',5'-monophosphate-dependent activation of mitogen-activated protein kinase in cumulus cells is essential for germinal vesicle breakdown of porcine cumulus-enclosed oocytes[J]. Endocrinology. 2005, 146(10): 4437-4444.
[19] Hansen D V, Pomerening J R, Summers M K, et al. Emi2 at the crossroads: where CSF meets MPF[J]. Cell Cycle. 2007, 6(6): 732-738.
[20] M O, D I, Kanemori Y E A. Erp1 /Emi2 is essential for the meiosis I to meiosis II transition in Xenopus oocytes[J]. Dev Biol. 2007, 303(1): 157-164.
[21] Ito J, Kato M, Hochi S, et al. Effect of enucleation on inactivation of cytostatic factor activity in matured rat oocytes[J]. Cloning Stem Cells. 2007, 9(2): 257-266.
[22] Gotze M, Kauffold P, Schuffenhauer A, et al. The inhibition of meiosis of bovine oocytes using biologic of synthetic inhibitors[J]. Arch Exp Veterinarmed. 1990, 44(1): 19-27.
[23]陈大元.受精生物学-受精机制与生殖工程[M].北京:科学出版社, 2000.
[24]王红铃.不同添加物对牛卵母细胞体外成熟、体外受精及受精卵体外培养的影响[D].南京农业大学, 2002.
[25] Gordon I. Laboratory production of cattle embryos[M]. UK: Commonwealth Agricultural Bureau International, Wallingford, Oxon, 1994.
[26] Jc J, Th C, Jk T, et al. Cytoskeletal patterns, in vitro maturation and parthenogenetic development of rabbit GV oocytes[J].Asian-Australasian Journal of Animal Sciences. 2002, 15(12): 1695-1701.
[27] Tao Y, Cao C, Zhang M, et al. Effects of cumulus cells on rabbit oocyte in vitro maturation.[J]. J Anim Physiol Anim Nutr (Berl). 2008, 92(4): 438-447.
[28] Wtk C, Rm M, C P. In vitro fertilization of pig and sheep oocytes matured in vivo[J]. Theriogenology. 1986, 25: 146.
[29] Lu K H, Gordon I, Gallagher M, et al. Pregnancy established in cattle by transfer of embryos derived from in vitro fertilisation of oocytes matured in vitro[J]. Vet Rec. 1987, 121(11): 259-260.
[30]郑瑞珍,高晓虹,王育哲,等.兔卵母细胞体外成熟和体外受精的研究[J].动物学报. 1992, 38(01): 66-72.
[31]赵晓娥,李向臣,高立功,等.山羊卵母细胞体外成熟研究[J].中国农学通报. 2005, 21(09): 1-4.
[32]索永善,李跃民,华云芳,等. PH值对猪卵母细胞体外成熟的影响[J].畜牧兽医杂志. 2005, 24(02): 4-5.
[33] Eppig J J. Maintance of meiotic arrest and the induction of oocyte maturation in mouse oocyte granulosa cell complexes developed in vitro from preantal follieles [J]. biol Reprod. 1991, 45: 824-830.
[34] Lu K H, Gordon I, Gallagher M, et al. Pregnancy established in cattle by transfer of embryos derived from in vitro fertilization of oocytes matured in vitro[J]. Vet Rec. 1987, 121(11): 259-260.
[35] Bd B, Ta R, T P. Development of in vitro matured in vitro fertilized bovine embryos into morulae and blastocysts in defined cu1ture media[J]. Theriogenology. 1992, 37(1): 127-146.
[36] Dag I, Ab S, Ec L. Deactivation of the serum prior to medium supplementation:a necessit or a myth[J]. Human Reprod. 1992, 7(supple2): 159.
[37]丁爱军,周艳华,贾银海,等.生殖激素对奶水牛卵母细胞成熟及发育潜能的影响[J].中国奶牛. 2008(11): 18-21.
[38]檀利长,田宁宁,霍道坦,等.牛卵泡液对牛卵母细胞体外成熟及受精胚发育力的影响[J].黑龙江动物繁殖. 2008, 16(05): 3-5.
[39] Ikeda S, Kitagawa M, Imai H, et al. The roles of vitamin A for cytoplasmic maturation of bovine oocytes[J]. J Reprod Dev. 2005, 51(1): 23-35.
[40] Dalvit G, Llanes S P, Descalzo A, et al. Effect of alpha-tocopherol and ascorbic acid on bovine oocyte in vitro maturation[J]. Reprod Domest Anim. 2005, 40(2): 93-97.
[41] Yanagimachi R. Gamete manipulation for development: new methods for conception[J]. Reprod Fertil Dev. 2001, 13(1): 3-14.
[42]冯伯森,王秋雨,胡玉兴编著.动物细胞工程原理与实践[M].北京:科学出版社, 2000.
[43]张红卫主编.发育生物学[M].北京:高等教育出版社, 2001.
[44]郭志勤主编.家畜胚胎工程[M].北京:中国科学技术出版社, 1998.
[45]秦鹏春,石惠芝,郭立民.试管牛胚胎的实验室生产过程[J]. 1997.
[46] Totey S M, Pawshe C H, Singh G P. Effects of bull and heparin and sperm concentrations on in vitro fertilization of buffalo (Bubalus bubalis ) oocytes matured in vitro[J]. Theriogenology. 1993, 39(4): 887-898.
[47] Parrish J J, Susko-Parrish J, Winer M A, et al. Capacitation of bovine sperm by heparin[J]. Biol Reprod. 1988, 38(5): 1171-1180.
[48]谭世俭,卢克焕.受精用液种类对牛卵母细胞体外受精的影响[J].中国兽医学报. 1994, 14(02): 140-145.
[49]兰宗保,许惠艳,保若钢,等.精子浓度和授精前孵育时间、精卵共孵育时间对猪卵母细胞体外受精效果的影响[J].广西畜牧兽医.2008,24(3):139-142.
[50]刘健,刘建民,张嘉保.哺乳动物体外受精若干问题[J].中国兽医学报. 1992, 12(03): 307-314.
[51]李子义,周琪,邹啸环,等.家兔显微受精影响因素及显微受精胚胎超微结构[J].中国兽医学报. 1997,(06):593-599.
[52] Kimura Y, Yanagimachi R. Mouse oocytes injected with testicular spermatozoa or round spermatids can develop into normal offspring[J]. Development. 1995, 121(8): 2397-2405.
[53] Wei H, Fukui Y. Fertilizability of ovine, bovine or minke whale (Balaenoptera acutorostrata) spermatozoa intracytoplasmically injected into bovine oocytes[J]. Zygote. 2000, 8(3): 267-274.
[54] Palermo G, Joris H, Devroey P, et al. Pregnancies after intracytoplasmic injection of single spermatozoon into an oocyte[J]. Lancet. 1992, 340(8810): 17-18.
[55] Butler E A, Masson G M. Development of a successful ICSI programme without the use of PVP[J]. Hum Reprod. 1997, 12(5): 1115-1116.
[56] Balaban B, Lundin K, Morrell J M, et al. An alternative to PVP for slowing sperm prior to ICSI[J]. Hum Reprod. 2003, 18(9): 1887-1889.
[57] Goto K. Bovine microfertilization and embryo transfer[J]. Mol Reprod Dev. 1993, 36(2): 288-290.
[58]曾勇,胡晓东,刘洪君,等.卵母细胞浆内单精子注射后电激活的临床应用[J].中华妇产科杂志. 2004, 39(2): 120.
[59]鹿群,陈子江,高选,等.钙离子载体A23187联合嘌呤霉素激活行卵母细胞质内单精子注射后未受精卵母细胞的研究[J].中华妇产科杂志. 2006, 41(3): 182-185.
[60]张晓慧,曹云霞,章志国,等.两种人工激活剂对卵胞浆内单精子注射后未受精卵母细胞激活效果的比较[J].中国妇产科临床杂志. 2009, 10(3): 204-206.
[61] G B, Bracket. Analysis of factors involved in the in vitro Production of bovine embryos[J]. Theriogenology. 1993,39(1): 43-64.
[62] Telford N A, Watson A J, Schultz G A. Transition from maternal to embryonic control in early mammalian development: a comparison of several species[J]. Mol Reprod Dev. 1990, 26(1): 90-100.
[63] Reed M. L编郭年藩摘译.猪胚胎的体外培养[J].国外畜牧科技. 1992, 19(4): 16-18.
[64]曹明聚.绵羊胚胎体外培养条件的研究进展[J].国外畜牧科技. 1996, 23(05): 27-30.
[65]王红玲.不同添加物对牛卵母细胞体外成熟、体外受精、及受精卵体外培养的影响[D].江苏南京:南京农业大学, 2005.
[66] Neira J A, Tainturier D, Pen M A, et al. Effect of the association of IGF-I, IGF-II, bFGF, TGF-b1, GM-CSF,and LIF on the development of bovine embryos produced in vitro[J]. Theriogenology. 2010(73): 595-604.
[67]贾秀芬,周燕华,刘慧雯,等.三种抗氧化剂对小鼠体外受精胚胎发育的影响[J].中国组织化学与细胞化学杂志. 2009, 15(04): 70-73.
[68]张永华.抗氧化剂对小鼠1一细胞期胚胎体外发育的影响[D].吉林:延边大学, 2006.
[69]唐军旺.维生素C和E及血清种类对水牛体外受精早期胚胎发育与凋亡影响的研究[D].广西南宁:广西大学, 2008.
[70]王敏康,刘冀珑,李光鹏,等. M16添加硫磺酸和EDTA支持昆明白小鼠体外受精并发育至囊胚[J].遗传. 2000, 22(05): 301-302.
[71]卢晟盛,魏凤,房慧伶,等.乙二胺四乙酸钠对猪卵母细胞的体外成熟和孤雌激活后早期发育的影响[J].安徽农业大学学报. 2008, 35(01): 9-16.
[72] Lu J H, Wang J Z, Wang H L, et al. Damaging effect of cumulus denudation on rabbit oocytes[J]. Fertil Steril. 2010, 93(5): 1567-1573.
[73]虹卢,杨蓉生,马志杰,等.牛卵母细胞体外成熟的研究进展[J].青海畜牧兽医杂志. 2006, 36(5): 44-45.
[74]戴荣亮,周俊波,王杏龙.山羊卵母细胞体外成熟的研究[J].中国草食动物. 2006, 26(1): 12-14.
[75] Al V S A E. FSH and growth factors affect the growth and endocrine function in vitro of granulose cells of bovine preantral follicles[J]. Theriogenology. 1996(45): 817-832.
[76]刘红林,范必勤.哺乳动物卵母细胞的体外成熟[J].畜牧与兽医. 1996, 28(4): 180-183.
[77]张德福,王建荣.兔卵母细胞体外培养系统的建立[J].上海农业学报. 1994, 10(03): 83-89.
[78]张德福,王济容.家兔卵巢卵母细胞体外成熟和体外受精的初步试验[J].上海农学院学报. 1993, 11(1): 12-16.
[79] L L P, J I. Enhancement of cumulus expansion and unclear maturation during bovine occyte maturation on in vitro by the addition of epidermal growth factor and insulin like growth factor[J]. Fertil, J Rep Rod. 1994, 101(3): 697-701.
[80] Wangw, K N. Syenergetic efficets of EGF factor and gonadotropins on the cytoplasmicmaturation of pig ooctyes in a serum-free medium[J]. Zygot. 1995(3): 345-350.
[81] Kw P. Exposure of bovine oocyte to EGF duringmaturation allows them to develop to blastcysts in a chemically - defined medium[J]. Theriogenology. 1997(48): 1127-1135.
[82]张涌,刘泽隆,李裕.山羊卵泡卵母细胞体外成熟的研究[J].西北农业大学学报. 1999,27(1): 14-18.
[83]冯贵雪,卞桂华,王晓丽,等.表皮生长因子对水牛卵母细胞体外培养核质成熟的影响[J].中国畜牧兽医. 2007, 34(1): 75-78.
[84]贺奋义,桑国俊,余四九,等.牛卵母细胞体外成熟技术研究[J].畜牧兽医杂志. 2008, 27(06): 16-18.
[85]进鲁,刘凤军,王勇胜,等. EGF和IGF - 1对山羊卵母细胞体外成熟的影响[J].黑龙江畜牧兽医. 2005(3): 7-9.
[86]马红,郭镇华,李忠秋,等.表皮生长因子在绵羊卵母细胞体外成熟过程中的作用[J].黑龙江畜牧兽医. 2007(7): 51-52.
[87] Lorenzo P L, Illera M J, Illera J C, et al. Enhancement of cumulus expansion and nuclear maturation during bovine oocyte maturation in vitro by the addition of epidermal growth factor and insulin-like growth factor I[J]. J Reprod Fertil. 1994, 101(3): 697-701.
[88] Grupen C G, Nagashima H, Nottle M B. Role of epidermal growth factor and insulin-like growth factor-I on porcine oocyte maturation and embryonic development in vitro[J]. Reprod Fertil Dev. 1997, 9(6): 571-575.
[89] Smitz J, Cortvrindt R, Hu Y. Epidermal growth factor combined with recombinant human chorionic gonadotrophin improves meiotic progression in mouse follicle-enclosed oocyte culture[J]. Hum Reprod. 1998, 13(3): 664-669.
[90] Lorenzo P L, Liu I K, Carneiro G F, et al. Equine oocyte maturation with epidermal growth factor[J]. Equine Vet J. 2002, 34(4): 378-382.
[91] Purohit G N, Brady M S, Sharma S S. Influence of epidermal growth factor and insulin-like growth factor 1 on nuclear maturation and fertilization of buffalo cumulus oocyte complexes in serum free media and their subsequent development in vitro[J]. Anim Reprod Sci. 2005, 87(3-4): 229-239.
[92] Lorenzo P L, Rebollar P G, Illera M J, et al. Stimulatory effect of insulin-like growth factor I and epidermal growth factor on the maturation of rabbit oocytes in vitro[J]. J Reprod Fertil. 1996, 107(1): 109-117.
[93] Gomez E, de Los S M, Ruiz A, et al. Effects of epidermal growth factor in the final stages of nuclear and cytoplasmic oocyte maturation in humans[J]. Hum Reprod. 1993, 8(5): 691-694.
[94]王炼炼,彦丘,幸贵邦.表皮生长因子促进小鼠未成熟卵母细胞体外成熟的实验研究[J].第三军医大学学报. , 29(22): 2179-2181.
[95]胥焘,窦忠英,陈静波. EGF和GSH对牛卵母细胞体外受精的影响[J].中国农业通报. 2005, 21(6): 55-57.
[96]赵振华,钱红娟,周敏敏,等.细胞因子与激素对山羊卵母细胞体外成熟及卵裂的影响[J].畜牧与兽医. 2008, 40(09): 38-40.
[97] Rodriguez A, De Frutos C, Diez C, et al. Effects of human versus mouse leukemia inhibitory factor on the in vitro development of bovine embryos[J]. Theriogenology. 2007, 67(5): 1092-1095.
[98]周敏敏,丁海雷,钱红娟,等. LIF对小鼠卵母细胞体外成熟和体外受精效果的影响[J].上海畜牧兽医通讯. 2008(03): 38-39.
[99]纪红,刘慧雯,秦逸人,等.显微操作针制作相关问题的探讨[J].生殖医学杂志. 2007, 16(6): 419-422.
[100] Nagy编孙青原陈大元主译.小鼠胚胎操作实验手册[Z].化学工业出版社, 2006.
[101]李嫒.人类辅助生殖实验技术[M].北京:科学出版社, 2008.
[102]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(04): 49-51.
[103]范必勤,熊慧卿,刘铁铮,等.家兔体外受精研究[J].江苏农业学报. 1987, 3(01): 11-17.
[104]任克良,罗惠娣,毛杨毅,等.家兔体外受精技术研究[J].华北农学报. 2000, 15(01): 135-138.
[105]蒋涛,高庆华,何良军,等.兔体外受精的研究[J].中国草食动物. 2005, 25(06): 3-6.
[106] Brackett B G, Mills J A, Jr Jeitles G G. In vitro fertilization of rabbit ova recovered from ovarian follicles[J]. Fertil Steril. 1972, 23(12): 898-909.
[107] Fukuda A, Roudebush W E, Thatcher S S. Platelet activating factor enhances the acrosome reaction, fertilization in vitro by subzonal sperm injection and resulting embryonic development in the rabbit[J]. Hum Reprod. 1994, 9(1): 94-99.
[108]周曾娣,黎曼侬,陈云鹤,等.超声断尾精子头显微受精-输卵管内移植研究的初步报告[J].中国优生与遗传杂志. 2003, 11(01): 72-73.
[109]陈浩杰,刘丽均,张春燕,等.实验兔胞内单精子注射技术[J].中国实验动物学报. 2008, 16(04): 250-253.
[110]唐修君.家兔超数排卵和体外受精技术的研究[D].江苏,扬州:扬州大学, 2007.
[111]李善刚,陈学进,李宏滨,等.不同激活条件和培养基对兔卵母细胞孤雌激活和胚胎发育的影响[J].畜牧兽医学报. 2008, 39(12): 1665-1670.
[112] Wang X, Falcone T, Attaran M, et al. Vitamin C and vitamin E supplementation reduce oxidative stress-induced embryo toxicity and improve the blastocyst development rate[J]. Fertil Steril. 2002, 78(6): 1272-1277.
[113]贾秀芬,郭铁云,周燕华,等.抗氧化剂对小鼠体外受精胚胎发育的影响[J].解剖科学进展. 2009, 15(1): 70-73.
[114]尚江华,张秀芳,黄右军.添加β-巯基乙醇对水牛胚胎体外发育的影响[J].草食家畜. 2001(03): 28-31.
[115] Park E S, Hwang W S, Kang S K, et al. Improved embryo development with decreased apoptosis in blastomeres after the treatment of cloned bovine embryos with beta-mercaptoethanol and hemoglobin[J]. Mol Reprod Dev. 2004, 67(2): 200-206.
[116] Goncalves F S, Barretto L S, Arruda R P, et al. Effect of antioxidants during bovine in vitro fertilization procedures on spermatozoa and embryo development [J]. Reprod Domest Anim. 2010, 45(1): 129-135.