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1.
猪ICM注入囊胚获得嵌合胚的研究 总被引：1，自引：0，他引：1
以湖北白猪囊胚为ＩＣＭ供体，杜洛克猪囊胚为受体囊胚，进行嵌合胚的研究，采用免疫外科和显微外科两种方法，分离１８４枚供体囊胚的ＩＣＭ。用酶将ＩＣＭ分散，而后注入到受体囊胚的囊胚腔中，共获６７枚嵌合胚，经体外短期培养，其中３５枚嵌合胚恢复，恢复率为５２．２％，将恢复的嵌合胚分别移入５头与供体囊胚猪同步的受体母猪子宫内，两头受孕，其中一头维持到分娩，产仔４头，但未见毛色嵌合现象。 相似文献
2.
采用核移植技术将单个细胞注入去核2-细胞卵裂球中,比较来自于囊胚的内细胞团(ICM)和滋胚层细胞(TE)产生孕体或嵌合体的发育潜力。用TE和ICM重构胚胎的发育潜力,主要取决于注射hCG后从输卵管收集到的2-细胞胚胎的时间。在注射hCG后38～42 h和43～46 h收集得到的2-细胞胚胎,其重构胚胎仅仅能够发育到4-细胞期。注射hCG后48～51 h收集得到的2-细胞胚胎,重构后发生卵裂的胚胎比率显著提高,有少部分会发育到囊胚期。用诺考达唑(Nocodazole)处理这些重构胚,发生卵裂的胚胎比率会显著提高,并且有部分重构胚发育到囊胚阶段。将这些囊胚移植到受体鼠中,在其妊娠中期未检测到供体核的出现。用ICM和TE进行重构得到的嵌合2-细胞胚胎,其体外发育潜力有限,本试验中也未得到嵌合孕体。 相似文献
3.
采用注入嵌合法初步建立了一套黄牛和水牛种间嵌合的程序与方法。采用机械剥离法或免疫外科法分离胚胎内细胞团(ICM),然后注入到已去除ICM的受体囊胚中构建形成水牛和黄牛的嵌合胚。结果发现,在用免疫外科法分离ICM时,抗血清的灭活温度从57℃升至63.5℃,ICM的获得数显著升高(0%vs100%,P<0.01),如若在分离培养液中添加6%的胎牛血清(FCS),ICM的获得数大大降低(97.6%vs0%,P<0.01)。采用免疫外科法分离得到的黄牛ICM进行水牛囊胚的ICM置换重组,重组胚的存活率与机械剥离法得到的ICM无显著差异(91.4%vs87.5%,P>0.05);但囊胚孵化率则显著提高(80%vs43.8%;P<0.05)。以上结果表明,⑴水牛和黄牛胚胎通过ICM置换获得的种间嵌合胚胎能继续发育;⑵用于黄牛ICM分离的兔抗牛抗血清需在63.5℃灭活30min,且分离需在无血清的培养液中进行;⑶通过分离ICM置换进行胚胎嵌合时,免疫外科法优于显微手术法。 相似文献
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梅花鹿新鲜和冷冻胚胎移植技术的研究 总被引：1，自引：0，他引：1
实验对12头供体母鹿进行超数排卵,27头受体同期发情后进行胚胎移植,建立梅花鹿胚胎移植技术体系。同期发情经埋置CIDR处理,超数排卵的供体每头分8次注射总剂量320 mg的FSH。供体发情鉴定后8~12 h通过直肠把握法进行人工授精。结果表明:冲胚时的胚胎回收率为74.39%(61/82),所获胚胎中,囊胚、桑椹胚、2-细胞、未受精卵所占的比例分别为1.64%(1/61)、77.05%(49/61)和1.64%(1/61)、19.67%(12/61)。研究将20枚新鲜胚胎移植给14头受体;将28枚胚胎经细管法玻璃化冷冻、解冻后移植给13头受体。鲜胚移植的受体妊娠率为85.71%(12/14),产仔率为64.29%(9/14);冻胚移植的受体妊娠率为61.54%(8/13),产仔率为53.85%(7/13),二者没有显著性差异(P0.05)。 相似文献
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兔核移植胚胎的克隆研究 总被引：12，自引：0，他引：12
本研究改进了兔受体卵母细胞的去核程序及供体卵裂球的处理方法，并对核移植胚的体外克隆、继代克隆及克隆胚的体内发育进行了试验。结果表明：卵龄对受体卵母细胞的去核具有显著的影响，卵龄为１５～１８ｈ的去核率为１００％（４５／４５），显著高于１３～１４ｈ的４６．６％（７／１５），Ｐ＜０．０１。ＤＮＡ合成抑制剂Ａｐｈｉｄｉｃｏｌｉｎ处理１６－细胞期卵裂球后，重组胚的囊胚发育率５４％（２０／３７）高于未处理组的４５％（１４／３１），Ｐ＜０．０５。从２枚１６－细胞供体胚分别获得来自一个供体胚的９枚和７枚核移植克隆囊胚，囊胚发育率为５１．６（１６／３１）。然而第一次继代核移植胚的囊胚发育率仅为１１．５％（６／５２）。１６－及３２－细胞期卵裂球的重组胚可发育至产仔，共获２窝８只仔兔。其中１只、２只、２只和３只仔兔分别来自４个供体胚。 相似文献
8.
对解冻后品质优良的Ａ级囊胚进行分割，并将二分胚移植生产同卵双生犊牛。结果表明：９枚Ａ级囊胚切割成１８枚半胚，胚胎分割成功率１００％。单胚移植受体６头。妊娠２头，妊娠率３３．３％；双半胚移植６头，妊娠２头，妊娠率３３．３％。 相似文献
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用简易分割方法,分割奶牛冷冻胚胎16枚,获得半胚32枚,分割成功率为100％(16/16),可移植半胚29枚,移植于16头受体牛,到第3个情期未返情者经直肠检查有6头妊娠,妊娠率为37.5％(6/16)。其中,在分割前或者分割后经恢复培养0.5～2h再移植的10头受体牛,5头妊娠;而未经恢复培养,分割后直接移植的6头受体牛中,只有1头妊娠。移植后3个月,直肠检查确定2头流产。已有1头受体黄牛生出1头奶牛牛犊(母)和1头受体奶牛产1头奶牛牛犊。其余的2头妊娠受体牛将于9月份产犊。此外,用简易分割法分割奶牛胚胎5枚,得到半胚10枚,裸半胚直接冷冻,解冻后回收可移植半胚5枚,移植于4头受体牛,无一头妊娠。结果表明,冷冻胚胎的分割半胚优于分割后冷冻半胚移植效果;冷冻胚胎分割前或者分割后恢复培养移植优于未经恢复培养而直接移植;简易分割法可应用于冷冻胚胎的分割。 相似文献
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Production of quail-chick chimaeras by blastoderm cell transfer 总被引：2，自引：0，他引：2
1. Quail-chick chimaeras were produced by injecting dissociated quail blastoderm cells into chick embryos. 2. Quail blastoderms were removed from the yolk and the cells were dispersed by trypsin treatment or pipetting. The cell suspension (1 to 5 microliters) was injected into the subgerminal cavity of unincubated chick embryos. The chick embryos were then cultured in recipient eggshells. 3. Quail blastoderm cells injected into the chick embryos adhered to the chick embryonic cells. The rates of hatching were 8.6% (38 chicks from 441 eggs) and 40.3% (48 chicks from 119 eggs) when the volumes of the cell suspension injected were 3 to 5 microliters and 1 microliter, respectively. 4. Seven out of 86 hatched birds were clearly identified as being chimaeric because part of the feather colouring was of quail specificity. In addition to these chimaeric birds, there were 8 chimaeric embryos which died before hatching. The distribution patterns of the quail feathers were varied among the chimaeric birds and embryos. 5. This technique provides a basis for the investigation of chick embryo cryopreservation, genetic transformation and analysis of cell lineage of chickens. 相似文献
12.
为了更全面的对四倍体胚胎的发育能力进行评估,本研究利用原位末端标记(TUNEL)法检测了电融合生产的小鼠四倍体囊胚细胞凋亡情况,同时用实时定量PCR方法检测了促凋亡基因Bax和抑凋亡基因Bcl-2在四倍体囊胚中的表达。结果显示:四倍体与体外二倍体胚胎发育到囊胚的比率没有显著差别(P0.05);四倍体囊胚无论是内细胞团细胞数、滋养层细胞数还是内细胞团/总细胞数比率都显著低于体内与体外二倍体囊胚(P0.05);四倍体囊胚细胞凋亡指数显著高于体内与体外二倍体囊胚细胞凋亡指数(P0.05);促凋亡基因Bax在四倍体囊胚中的表达显著高于体内与体外二倍体囊胚中的表达(P0.05),而抑凋亡基因Bcl-2在四倍体囊胚中的表达同体内与体外二倍体囊胚中的表达没有显著差异(P0.05)。因此,四倍体囊胚质量比体内与体外二倍体囊胚质量都差,这可能是四倍体胚胎不能发育到足月的原因之一。 相似文献
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Production of ovine chimeras by inner cell mass transplantation 总被引：2，自引：0，他引：2
J E Butler G B Anderson R H BonDurant R L Pashen M C Penedo 《Journal of animal science》1987,65(1):317-324
Ovine chimeras were produced by micro-injection of isolated inner cell masses (ICM) into recipient blastocysts. Inner cell masses were isolated by immunosurgery. A total of 57 chimeric embryos was produced, 52 of which were transferred to recipient ewes. Thirty-seven live lambs were born, of which 15 were determined to be chimeric on the basis of blood type analysis. One lamb, although not a blood chimera, exhibited overt signs of chimerism. An additional six lambs were determined to have developed solely from the injected ICM. The rate of chimerism in live lambs was 43% (16/37) while the survival rate of injected ICM was 59% (22/37). The method presented allows the production of relatively large proportion of viable, chimeric embryos without the use of an intermediate recipient. 相似文献
15.
Inhalt: Rinderchimären wurden durch Aggregation geteilter Embryonen unterschiedlicher Rassen im 16- bis 32-Blastomerenstadium erstellt. Nach Transfer von insgesamt 25 chimären Embryonen wurden neun (36%) Kälber geboren, von denen vier = 44,4% chimär waren. Der Nachweis des Chimärismus erfolgte durch Blutgruppenfaktoren, Chromosomenstruktur, Fellpigmentierung und Rückkreuzungen. Im einzelnen konnten durch die Blutgruppenfaktoren zwei = 22,2%, über die Chromosomenstruktur ein = 11,1% und durch die Fellfarbe drei = 33,3% Tiere als chimär erkannt werden. Zwei Kälber, die in ihrem äuβeren Erscheinungsbild die Ausgangsrassen Angler und Schwarzbunt repräsentierten, besaßen identische Blutgruppen. Durch Rückkreuzungen mit zwei chimären Bullen wurde erstmals ein Chimärismus der Keimzellen nachgewiesen .
Contents: Investigations on the production and genetic characterisation of chimaeric cattle
Cattle chimaeras were produced by aggregation of split embryos from different breeds between 16- to 32-cell stage. Altogether 9 (36%) out of 25 transferred chimaeric embryos developed to term of which four proved to be chimaeric. In order to confirm chimaerism blood groups, chromosome structure and coat colour pigmentation were determined and back-crossings were performed. Evidence for chimaerism was given by blood groups (2 = 22,2%), chromosome structure (1 = 11,1%) and coat colour pigmentation (3 = 33,3%). Two calves showing different coat colour pigmentation patterns of either the Angler or the Frisian appeared to have identical chimaeric blood group compositions. By back-crossings of two chimaeric bulls germ cell chimaerism could be demonstrated for the first time . 相似文献
Contents: Investigations on the production and genetic characterisation of chimaeric cattle
Cattle chimaeras were produced by aggregation of split embryos from different breeds between 16- to 32-cell stage. Altogether 9 (36%) out of 25 transferred chimaeric embryos developed to term of which four proved to be chimaeric. In order to confirm chimaerism blood groups, chromosome structure and coat colour pigmentation were determined and back-crossings were performed. Evidence for chimaerism was given by blood groups (2 = 22,2%), chromosome structure (1 = 11,1%) and coat colour pigmentation (3 = 33,3%). Two calves showing different coat colour pigmentation patterns of either the Angler or the Frisian appeared to have identical chimaeric blood group compositions. By back-crossings of two chimaeric bulls germ cell chimaerism could be demonstrated for the first time . 相似文献
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Takashi FUJII Nobuyuki SAKURAI Tsubasa OSAKI Gentaro IWAGAMI Hiroki HIRAYAMA Akira MINAMIHASHI Tsutomu HASHIZUME Ken SAWAI 《The Journal of reproduction and development》2013,59(2):151-158
In mouse embryos, segregation of the inner cell mass (ICM) and trophectoderm (TE)
lineages is regulated by genes, such as OCT-4, CDX2 and
TEAD4. However, the molecular mechanisms that regulate the segregation
of the ICM and TE lineages in porcine embryos remain unknown. To obtain insights regarding
the segregation of the ICM and TE lineages in porcine embryos, we examined the mRNA
expression patterns of candidate genes, OCT-4, CDX2,
TEAD4, GATA3, NANOG,
FGF4, FGFR1-IIIc and FGFR2-IIIc, in
blastocyst and elongated stage embryos. In blastocyst embryos, the expression levels of
OCT-4, FGF4 and FGFR1-IIIc were
significantly higher in the ICM than in the TE, while the CDX2,
TEAD4 and GATA3 levels did not differ between the ICM
and TE. The expression ratio of CDX2 to OCT-4
(CDX2/OCT-4) also did not differ between the ICM and
TE at the blastocyst stage. In elongated embryos, OCT-4,
NANOG, FGF4 and FGFR1-IIIc were
abundantly expressed in the embryo disc (ED; ICM lineage), but their expression levels
were very low in the TE. In contrast, the CDX2, TEAD4
and GATA3 levels were significantly higher in the TE than in the ED. In
addition, the CDX2/OCT-4 ratio was markedly higher in
the TE than in the ED. We demonstrated that differences in the expression levels of
OCT-4, CDX2, TEAD4,
GATA3, NANOG, FGF4,
FGFR1-IIIc and FGFR2-IIIc genes between ICM and TE
lineages cells become more clear during development from porcine blastocyst to elongated
embryos, which indicates the possibility that in porcine embryos, functions of ICM and TE
lineage cells depend on these gene expressions proceed as transition from blastocyst to
elongated stage. 相似文献
17.
Thanh Quang Dang‐Nguyen Nguyen Viet‐Linh Tamas Somfai Seiki Haraguchi Bui Xuan Nguyen 《Animal Science Journal》2014,85(5):517-523
In the present study, we examined the development to blastocysts of large and small blastomeres from unevenly cleaved 2‐cell embryos (uneven 2‐cell embryos) in pigs. Proportion of blastocysts derived from large blastomeres (52.8 ± 6.4%) was significantly higher (P < 0.05) compared with small ones (32.1 ± 4.6%). However, there were no differences in total cell number, inner cell mass (ICM) cell number and ICM/total cells ratio between them. Of 53 sister blastomere pairs in the same embryos examined there were 12 pairs (22.6%) in which both blastomeres developed to blastocysts, 16 pairs (30.2%) in which only large blastomeres developed to blastocysts, and five pairs (9.4%) in which only small blastomeres developed to blastocysts. Relative total amount of active mitochondria in small blastomeres were lower (P < 0.05) than that of large blastomeres and blastomeres from evenly cleaved 2‐cell embryos. However, there was no difference in relative density of active mitochondria in these three types of blastomeres. In conclusion, blastocysts derived from small and large blastomeres in uneven 2‐cell embryos had comparable quality in terms of cell number, ICM number, ICM/total cell ratio and distribution of active mitochondria. The results suggest that these blastomeres may contribute multiple offspring production in pigs. 相似文献
18.
Ambikaprasanna Saha Manmohan S. Chauhan Radhey S. Manik Prabhat Palta Suresh K. Singla 《Reproduction in domestic animals》2023,58(1):158-167
In this study we treated the handmade cloned (HMC) buffalo embryos with the DNA methylation inhibitors; 5-aza-2′-deoxycytidine (AzadC) or Zebularine individually after post-fusion and during in vitro culture till eighth day. The blastocysts production rate significantly improved (p < .01) after treating embryos independently with 5 nM AzadC and 5 nM zebularine compared with 2 and 10 nM AzadC or zebularine groups, respectively. The highest cleavage rates were obtained for 5 nM treatment of AzadC and zebularine compared with other treatments and untreated control group. Quality of blastocysts were evaluated using total cell number (TCN) and the ratio of number of inner cell mass (ICM) cells/total cell number (ICM/TCN). Zebularine treatments (2/5/10 nM) significantly improved both TCN and ICM/TCN ratio compared with AzadC treatments (2/5/10 nM); however, control group TCN and ICM/TCN ratio was found lower. The methylation percentage of pDS4.1 and B. bubalis satellite DNA were comparatively more attenuated with 5 nM zebularine than 5 nM AzadC treatment. The increased in vitro development rates of the treated embryos were correlated with the decreased level of DNA methylation and the improved blastocyst quality. Following transfer of 5 nM zebularine treated embryos to 6 recipients, 4 were found to be pregnant, though the pregnancies were not carried to full term. 相似文献