共查询到20条相似文献，搜索用时 187 毫秒
1.
扩张囊胚玻璃化超低温冷冻保存及移植技术的研究 总被引：8，自引：0，他引：8
本试验探讨了对鼠扩张囊胚玻璃化冷冻保存后，提高发育率的最佳条件。玻璃化溶液是用ＰＢＳ液稀释成的３０％聚蔗糖＋０．５Ｍ蔗糖混合法，再将乙二醇（ＥＧ）调制成２０、３０及４０％（ｖ／ｖ）的溶液，即ＥＦＳ２０、３０及４０。玻璃化冷冻保存是１０、２０及２５℃温度下，将扩张囊胚直接移入ＥＰＳ溶液后投入液氮中一步法令冻保存，其中２５℃下保存后的发育率最高（８７％）。继之将扩张囊胚用１０％ＥＧ溶液经５分钟处理后，再移入ＥＦＳ４０中二步法冷冻保存，发育率高达９４％。利用二步法冷冻保存的扩张囊胚移植后，妊娠受体的产仔率为６６％（１６８／２５５），同时照组产仔率６１％（８０／１３２）相比无显著性差异（Ｐ＞０．０５）。 相似文献
2.
生物频谱对家兔胚胎发育影响的初步研究 总被引：4，自引：1，他引：3
本文首次研究了周林生物频谱照射对家兔胚胎体外发育的影响，初步探讨了周林生物频谱在胚胎工程中应用的可能性。周林生物频谱分别照射家兔２－细胞胚胎３、５、７、１０、１５、２０、２５分钟，结果照射１０、１５、２０、２５分钟，可使７２小时（从注射ＨＣＧ后算起，即照射后４８小时）桑椹胚发育率显著增加，照射２０和２５分钟可提高７２、８４、９６和１０８小时囊胚发育率，照射１０分钟和１５分钟可提高１０８小时囊胚发育率，照射７、１０、１５、２０和２５分钟可提高９６小时扩张囊胚发育率及１０８小时孵化囊胚发育率。周林生物频谱照射家兔１６－细胞胚胎２０分钟，其６０小时（从注射ＨＣＧ后起，即照射后１２小时）致密桑椹胚发育率与对照比较差异极显著（Ｐ＜０．０１），囊胚、扩张囊胚和孵化囊胚发育率与对照比较无显著差异（Ｐ＞０．０５） 相似文献
3.
家兔扩张囊胚玻璃化冷冻保存技术的研究 总被引：16，自引：0，他引：16
本试验对家兔扩张囊胚的玻璃化冷冻保存技术进行了探讨。首先在２０℃室温下，将扩张囊胚直接移入ＥＦＳ４０溶液中短时间平衡后，直接投入液氮中冷冻保存（一步法）；或胚胎在１０％～２０％ＥＧ（或ＥＦＳ２０）溶液中经预先处理后，再移入ＥＦＳ４０中冷冻保存（二步法），解冻后的胚胎发育率达到９５％～１００％。当室温提高到２５℃时，一步法和二步法冷冻的胚胎均得到了较高的发育率（８９％～９０％）。利用２０℃条件下一步法即２分钟平衡后冷冻的胚胎移植后，妊娠受体的产仔率为２９．２％（７／２４），同对照组产仔率３２．４％（１１／３４）相比无显著性差异（Ｐ＞０．０５）。 相似文献
4.
5.
以猪孤雌激活胚胎为材料，经体外成熟、电激活，选取电激活后3d（6--8cell）胚胎分别放入0．28mol／L的蔗糖离心液或无蔗糖离心液中离心处理后，继续培养5d，取扩张囊胚进行玻璃化冷冻保存。结果显示，在囊胚形成率上，无蔗糖组（19．86％）略高于蔗糖组（18．83％），差异不显著（P〉0．05）；胚胎复苏率上两者差异不显著，但蔗糖组（43．33％）明显好于无蔗糖组（29．63％）；在解冻后胚胎内细胞破损率上，无蔗糖组（20．18％）低于蔗糖组（29．13％），差异不显著（P〉0．05）。因此，离心液中添加蔗糖，对后续囊胚形成无影响，在一定程度上提高了解冻囊胚复苏率，但没有降低内细胞的破损程度。 相似文献
6.
小鼠桑椹胚简易玻璃化冷冻技术再探讨 总被引：12，自引：0，他引：12
本试验继小鼠扩张囊胚玻璃化冷冻保存成功后，在室温（２５℃）下利用不同浓度的ＥＦＳ玻璃化溶液，对小鼠的桑椹胚简易玻璃化冷冻技术进行再探讨。结果是胚胎在１０％ＥＧ溶液中预先处理５分钟，再移入事先配置好含有ＥＦＳ３０的０．２５ｍｌ塑料细管中１分钟平衡后直接投入液氮中冷冻，解冻后获得的发育率最高（９４％）。冻胚移植后妊娠率和产仔率分别为５６％（９／１６）及４２％（４９／１１６）。与对照组相比差异不显著（Ｐ＞０．０５） 相似文献
7.
绵羊冷冻胚胎有枚浆冻前Ａ级胚胎解冻后,胚胎等级有所下降,可用胚率为７７．９％,Ａ、Ｂ级胚移植妊娠率分别为３３．３％（１８／５４）和１６．７％（１／６）,差异不显著（Ｐ〉０．０５）;桑椹胚、早期囊胚、中期囊胚、扩张囊胚移植妊娠率分别为２０．８％（５／２５）、４２．９％（６／１４）、３５．７％（５／１４）５ ３７．５％（３／８）,囊胚期胚胎移植妊娠率较高,但差异不显著（Ｐ〉０．０５） 相似文献
8.
牛体外受精胚冷冻保存的研究 总被引：9，自引：0，他引：9
对牛卵泡卵母细胞体外受精（ＩＶＦ）１６８ｈ的致密桑椹胚、囊胚用常规快速冷冻法、预冷和不预冷的超快速冷冻法进行了冷冻保存试验。结果表明：ＩＶＦ囊胚采用含１０％甘油的常规快速冷冻法、含２．１ｍｏｌ／Ｌ甘油和０．２５ｍｏｌ／Ｌ蔗糖预冷５ｍｉｎ的一步冷冻法及含２５％甘油和２５％乙二醇预冷５ｍｉｎ的玻璃化冷冻法等３种方法进行冷冻保存，解冻后的继续发育率（６８．０％，５９．０％，６５．７％）均无显著差异（Ｐ＞０．０５），可用快速、简便、预冷的一步冷冻法或玻璃化冷冻法替代常规快速冷冻法；ＩＶＦ致密桑椹胚可用一步冷冻法（２．１ｍｏｌ／Ｌ甘油＋０．２５ｍｏｌ／Ｌ蔗糖）和玻璃化冷冻法（２５％甘油＋２５％乙二醇或２５％甘油＋２５％１，２－丙二醇）进行预冷的超快速冷冻保存；冻前预冷（５ｍｉｎ）能显著提高ＩＶＦ囊胚的冻后形态正常率和继续发育率（Ｐ＜０．０５）；ＩＶＦ囊胚冷冻—解冻后的继续发育率高于ＩＶＦ致密桑椹胚。 相似文献
9.
脱防冻剂方法对牛体外受精胚胎冷冻后成活率的影响 总被引：10，自引：0，他引：10
为了观察脱防冻剂方法对牛体外受精冷冻胚胎在体内、外发育率的影响,应用常规冷冻法在０．７５ｍｏｌ／Ｌ甘油＋０．５ｍｏｌ／Ｌ丙二醇溶液中冷冻受精后７、８ｄ的囊胚,解冻后在０．２５、０．５ｍｏｌ／Ｌ蔗糖液中二步或在０．２５ｍｏｌ／Ｌ蔗糖液中三步脱防冻剂的胚胎孵化率（分别为６８．６％、６２．２％、６８．７％）与用ＰＢＳ／ＦＣＳ六步脱防冻剂的差异不显著（７０．４％,Ｐ＞０．０５）,但在０．５ｍｏｌ／Ｌ蔗糖液中三步脱防冻剂后,孵化率显著降低（４７．６％,Ｐ＜０．０１）。用０．２５ｍｏｌ／Ｌ或０．５ｍｏｌ／Ｌ蔗糖或海藻糖预先使胚胎脱水后冷冻,解冻后可使胚胎直接在ＰＢＳ中一步脱掉防冻剂,胚胎孵化率（４５．０％～４９．５％）与在０．２５ｍｏｌ／Ｌ蔗糖液中二步脱防冻剂的相似（５１．５％,Ｐ＞０．０５）。移植试验表明,在０．２５ｍｏｌ／Ｌ蔗糖液中二步脱防冻剂获得的妊娠率与六步脱防冻剂相似,与在ＰＢＳ中一步脱防冻剂的亦无明显差异,但妊娠率均偏低（２１．４％～３７．５％）。 相似文献
10.
生产条件下安哥拉山羊胚胎徒手分割研究 总被引：2，自引：0，他引：2
生产条件下,徒手二分割安哥拉山羊桑椹胚和囊胚,并将裸半胚手术移植于受体羊子宫角。１９９３年用０．５％链霉蛋白酶处理胚胎１～６ｍｉｎ软化透明带后,在玻璃培养皿中的含１２．５％蔗糖和５％新生犊牛血清（ＮＣＳ）的ＰＢＳ液内分割,将３３枚半胚移植到２１只受体,有２只受体妊娠。１９９４年用０．２５％链霉蛋白酶处理胚胎０．５～１ｍｉｎ软化透明带后或不经处理而直接在磨砂玻璃皿中的含２０％血清的ＰＢＳ液内分割,移植受体３４只,有１０只受体产羔（２９．４％）,共产羔１１只,其中１对为同卵双生,半胚成羔率为２０．０％（１１／５５）。１９９５年,囊胚不经处理直接在磨砂玻璃皿中的含２０％ＮＣＳ的ＰＢＳ液内分割,半胚成对移植于受体羊黄体侧子宫角,产羔受体率为５０．０％（１３／２６）,共产羔１６只,其中３对为同卵双生,半胚成羔率为３０．８％（１６／５２）,胚胎成羔率为６１．５％（１６／２６）。 相似文献
11.
Yang QE Hou YP Zhou GB Yang ZQ Zhu SE 《The Journal of reproduction and development》2007,53(2):211-218
In the present study, mouse blastocysts were employed to investigate the feasibility and efficiency of stepwise in-straw dilution and direct transfer using the open pulled straw (OPS) method. In experiment I, the effects of various vitrification solutions (VS) on embryo survival were examined. After thawing, the expanded blastocyst rates (97.59 and 95.05%) and hatching rates (80.48 and 78.95%) achieved in the EDFS30 [15% ethylene glycol (EG), 15% dimethyl sulfoxide (DMSO), Ficoll, and sucrose] and EFS40 [40% EG, Ficoll, and sucrose] groups were no different from those (96.15% and 83.33%) of the control group. However, the rates in the EFS30 [30% EG, Ficoll, and sucrose] (87.80 and 55.43%) and EDFS40 [20% EG, 20% DMSO, Ficoll, and sucrose] (95.69 and 70.97%) groups were significantly lower than those (96.15 and 83.33%) of the control group (P<0.05). In the experiment II, the effects of the volume of VS in the OPS on the survival of embryos after in-straw thawing were investigated. When the length of the VS in the column was less than 1 cm, the in vitro viability of embryos thawed by stepwise in-straw dilution was no different among the experimental and control groups. The embryos could be successfully thawed by immersing the OPS in 0.5 M sucrose for 3 min and then 0.25 M sucrose for 2 min. In experiment III, the effect of immersion time of the OPS in diluent (PBS) on the viability of vitrified embryos was investigated. After in-straw thawing, OPSs were immersed immediately in 1 ml PBS for 0 to 30 min. When the immersion time of the OPSs in PBS was less than 12 min, in vitro development of the in-straw thawed embryos was no different from that of the controls. In experiment IV, in-straw thawed blastocysts were directly transferred to pseudopregnant mice to examine their in vivo developmental viability. The pregnancy (91.67%) and birth rates (42.42%) of embryos in-straw thawed and directly transferred were no different from those of the unvitrified controls (90.90 and 40%) and embryos thawed by the conventional method (84.61 and 46.94%). These results demonstrate that mouse embryos vitrified with OPS could be successfully thawed by stepwise in-straw dilution and transferred directly to a recipient and that this method might be a model for field manipulation of vitrified embryos in farm animals. 相似文献
12.
牛体外受精胚胎一步脱防冻剂冷冻方法的研究 总被引：7，自引：1，他引：7
为了研究适合于牛体外受精胚胎的一步除防冻剂的冷冻方法，特进行两个试验。试验１分别用１．５Ｍ甘油＋０．２５Ｍ蔗糖、１．５Ｍ乙二醇和１．５Ｍ丙二醇作防冻剂冷冻体外受精后第７天，已发育至囊胚阶段的牛胚胎。使胚胎降温至－７℃后，植冰，然后以０．３℃／分的速率降温至－３０℃，立即将胚胎投入液氮中冷冻保存。在３７℃水中解冻胚胎后，使其直接在含１５％胎犊血清（ＦＣＳ）的磷酸缓冲液（ＰＢＳ）中脱去防冻剂。经体外培养７２ｈ后，３组胚胎的孵化率分别为７７．２７％（１０２／１３２）、７３．２４％（１０４／１４２）、４７．９０％（５７／１１９），第１、２组的孵化率极显著高于第３组（Ｐ＜０．０１），说明用１．５Ｍ丙二醇溶液冷冻的胚胎，在解冻后不宜直接在ＰＢＳ中脱防冻剂。试验２比较用不同浓度（１．０Ｍ０１．５Ｍ，２．０Ｍ）的乙二醇和不同投液氮温度（－２５℃，－３０℃，－３５℃）冷冻胚胎的效果。结果表明乙二醇浓度和投液氮温度对胚胎孵化率均无显著影响（Ｐ＞０．０５），各组胚胎的孵化率变动于７４．４４％－８５．４８％之间。 相似文献
13.
14.
Silva MA Peixoto GC Sousa PC Bezerra FS Bezerra AC Silva AR 《Reproduction in domestic animals》2012,47(1):e4-e6
This study verifies the interactions between straw size and thawing rates and their impact on the epididymal sperm from this species. Caudae epididymidum from 10 agoutis were subjected to retrograde washing using a coconut water extender (ACP‐109c®). Epididymal sperm were evaluated and extended in ACP‐109c® plus egg yolk (20%) and glycerol (6%). The samples were packaged in 0.25‐ or 0.50‐ml straws, frozen in liquid nitrogen and thawed at 37°C/1 min or 70°C/8 s, followed by a re‐evaluation. The use of 0.25‐ml straws thawed at 37°C/1 min provided a value of 26.6% for sperm motility. No interactions between straw size and thawing rates were verified on agouti sperm (p > 0.05), but when 0.5‐ml straws were thawed at 70°C/8 s, sperm vigour decreased significantly (p < 0.05). It is recommended that the agouti epididymal sperm cryopreserved in ACP‐109c® extender should be packaged in 0.25‐ or 0.50‐ml straws and thawed at 37°C/60 s. 相似文献
15.
In vitro development of OPU‐derived bovine embryos cultured either individually or in groups with the silk protein sericin and the viability of frozen‐thawed embryos after transfer 
下载免费PDF全文
下载免费PDF全文 Tomohiro Isobe Yoshihisa Ikebata Lanh Thi Kim Do Fuminori Tanihara Masayasu Taniguchi Takeshige Otoi 《Animal Science Journal》2015,86(7):661-665
The optimization of single‐embryo culture conditions is very important, particularly in the in vitro production of bovine embryos using the ovum pick‐up (OPU) procedure. The purpose of this study was to examine the development of embryos derived from oocytes obtained by OPU that were cultured either individually or in groups in medium supplemented with or without sericin and to investigate the viability of the frozen‐thawed embryos after a direct transfer. When two‐cell‐stage embryos were cultured either individually or in groups for 7 days in CR1aa medium supplemented with or without 0.5% sericin, the rates of development to blastocysts and freezable blastocysts were significantly lower for the embryos cultured individually without sericin than for the embryos cultured in groups with or without sericin. Moreover, the rate of development to freezable blastocysts of the embryos cultured individually with sericin was significantly higher than that of the embryos cultured without sericin. When the frozen‐thawed embryos were transferred directly to recipients, the rates of pregnancy, abortion, stillbirth and normal calving in the recipients were similar among the groups, irrespective of the culture conditions and sericin supplementation. Our findings indicate that supplementation with sericin during embryo culture improves the quality of the embryos cultured individually but not the viability of the frozen‐thawed embryos after transfer to recipients. 相似文献
16.
Embryos were flushed on day 7 after ovulation from two mares, and frozen using a conventional slow freezing procedure in phosphate buffered (PBS) saline supplemented with 10% FCS, 1.5 mol/L ethylene glycol and 0.25 mol/L sucrose. One of the two embryos was thawed after 10 months of storage in liquid nitrogen and transferred directly (without dilution of the cryoprotectant and quality examination) to a synchronized recipient. This transfer resulted in the birth of a live female foal. To our knowledge, this is the first live foal born after direct transfer of a frozen-thawed equine embryo. 相似文献
17.
U. Darvelid H. Gustafsson M. Shamsuddin B. Larsson H. Rodriguez Martinez 《Acta veterinaria Scandinavica》1994,35(4):417
The capacity of different vitrification media and methods was tested onto in vivo and in vitro produced bovine morula/blastocysts and their ultrastructure and survival studied post-thawing. Two vitrification solutions were finally selected, named 40 ES (40% ethylene glycol in PBS containing 0.5 M sucrose) and 35 EFS (composed of 35% (v/v) ethylene glycol in PBS containing 0.5 M/l sucrose and 30% (w/v) Ficoll 70). The straws were either precooled or not precooled in nitrogen vapour, plunged and stored in LN2 for 10–25 days, and then thawed in a 20° C waterbath. The content of the straws was rediluted in 1M sucrose solution in PBS and later cocultured with BOEC for 48 h. The overall survival rates for in vitro and in vivo embryos were 36% (12 of 33) and 20% (3 of 15) after 24 h and 21% (7 of 33) and 33% (5 of 15 ) after 48 h. The survival rates for precooled embryos were significantly higher than for not precooled (48% vs 13% after 24 h and 44% vs 4% after 48 h) when tested across vitrification media. The in vitro-produced embryos presented an ultrastructure similar to the pre-freeze state, irrespective of the vitrification media used. The in vivo developed embryos showed a rather modified post-thaw ultrastructure, with clear signs of osmotic changes at both the trophoblastic and embryonic cells. The results indicated that in vitro and in vivo developed bovine embryos can survive vitrification using ethylene glycol as a cryoprotectant. 相似文献
18.
Takeshi Hayashi Kazuki Kansaku Takahito Abe Shuji Ueda Hisataka Iwata 《Animal Science Journal》2019,90(7):849-856
This study evaluated the effects of cryopreservation by slow freezing on the mitochondrial function, DNA integrity, and developmental ability of bovine embryos and examined whether resveratrol treatment of the frozen‐thawed blastocysts improved embryonic viability. In vitro produced bovine embryos were subjected to slow freezing. After thawing, the ATP content and mitochondrial DNA integrity (mtDNA), determined by real‐time PCR targeting short and long mitochondrial sequences, was found to be lower in frozen‐thawed embryos than in fresh embryos, and mtDNA copy number was significantly reduced during the 24‐hr incubation post warming. Furthermore, immunostaining against double‐strand DNA revealed DNA damage in frozen‐thawed embryos. When frozen‐thawed embryos were incubated in the medium containing 0.5 µM resveratrol, SIRT1 expression, and survival rate of the embryos significantly improved compared with the vehicle‐treated embryos. In addition, cell‐free mtDNA content in medium was higher in case of resveratrol‐treated embryos than of vehicle‐treated embryos. In conclusion, slow freezing affects mitochondrial integrity and function in the blastocysts. In the frozen‐thawed embryos, mitochondria were removed during post‐thawing incubation and resveratrol enhanced the process, resulting in improved survivability of the embryos. 相似文献
19.
产蛋种鸡日粮中不同水平维生素E与有机和无机硒的效果研究 总被引：17，自引：2，他引：15
在３×５的两因子试验中,研究了不同添加水平维生素Ｅ（４、１４和２４ｍｇ／ｋｇ）与有机和无机硒（亚硒酸钠０．２０、０．３５和０．５０ｍｇ／ｋｇ,酵母硒０．３５和０．５０ｍｇ／ｋｇ）对褐壳蛋种母鸡在２０～４０周龄阶段生产繁殖性能和有关血液指标的影响。结果表明,产蛋母鸡的采食量、产蛋率、蛋重、产蛋量、饲料转化率和死淘率在３个维生素Ｅ添加水平间无差异;除了添加酵母硒０．５０ｍｇ／ｋｇ组蛋鸡的蛋重和产蛋量显著低于酵母硒０．３５ｍｇ／ｋｇ组以外,其它各项生产性能指标在各硒处理组间差异不明显。维生素Ｅ和硒可增加彼此在血液中的存留;当日粮中只补加维生素Ｅ４ｍｇ／ｋｇ时,将日粮亚硒酸钠硒添加水平由０．２０ｍｇ／ｋｇ提高到０．３５～０．５０ｍｇ／ｋｇ可使血浆α－生育酚含量由０．５０μｇ／ｍｌ上升到接近１．０μｇ／ｍｌ左右。 相似文献
20.
Hysteroscopic insemination of low numbers of flow sorted fresh and frozen/thawed stallion spermatozoa 总被引：4，自引：0，他引：4
The objective of this experiment was to determine the effects of flow cytometric sorting and freezing on stallion sperm fertility. A 2 x 2 factorial design was used to delineate effects of flow sorting and freezing spermatozoa. Oestrus was synchronised (July-August) in 41 mares by administering 10 ml altrenogest (2.2 mg/ml) per os for 10 consecutive days, followed by 250 microg cloprostenol i.m. on Day 11. Ovulation was induced by administering 3,000 iu hCG i.v. either 6 h (fresh spermatozoa) or 30 h (frozen/thawed spermatozoa) prior to insemination. Mares were assigned randomly to one of 4 sperm treatment groups. Semen was collected from 2 stallions with an artificial vagina and processed for each treatment. Treatment 1 (n = 10 mare cycles) consisted of fresh, nonsorted spermatozoa and Treatment 2 (n = 16 mare cycles) of fresh, flow sorted spermatozoa. Spermatozoa to be sorted were stained with Hoechst 33342 and sorted into X- and Y-chromosome-bearing populations based on DNA content using an SX MoFlo sperm sorter. Treatment 3 (n = 16 mare cycles) consisted of frozen/thawed nonsorted spermatozoa (frozen at 33.5 x 106 sperm/ml in 0.25 ml straws) and Treatment 4 (n = 15 mare cycles) of flow sorted frozen/thawed spermatozoa (frozen at 64.4 x 10(6) sperm/ml). Concentrations of sperm in both cryopreserved treatments were adjusted, based on predetermined average post-thaw motilities, so that each insemination contained approximately 5 x 10(6) motile spermatozoa. Hysteroscopic insemination of 5 x 10(6) motile spermatozoa in a volume of 230 microd was used for all treatments. Pregnancy was determined ultrasonographically 16 days postovulation. No differences were found (P>0.1) in the pregnancy rates for mares inseminated with fresh nonsorted (4/10 = 40.0%), fresh flow sorted (6/16 = 37.5%), frozen/thawed nonsorted (6/16 = 37.5%) and flow sorted frozen/thawed spermatozoa (2/15 = 133%). Pregnancy rates tended (P = 0.12) to be lower following insemination of frozen/thawed flow sorted spermatozoa. Further studies are needed with a larger number of mares to determine if fertility of flow sorted frozen/thawed spermatozoa can be improved. 相似文献