卵母细胞体外成熟时间对绵羊核移植效率的影响(英文)

[Objective] The study aimed to provide references for the time of oocyte maturation in vitro and enucleation in the course of sheep nuclear transfer(NT).[Method] Compared the effects of different maturation time of oocytes on enucleation efficiency and reconstructed embryo development by means of blind enucleation and fluorescence microscopy.[Result] Treatment of IVM(in vitro maturation)19-21 h was significantly higher than IVM 16-18 h treatment in oocyte maturation rate(P<0.05)and was significantly higher than IVM 22-24 h treatment in enucleation rate(P<0.05).Three treatments had no significant difference in cleavage rate and blastocyst rate(P>0.05),but IVM 19-21 h treatment was significantly higher than the other 2 treatments in average cell number of blastocysts(P<0.05).[Conclusion] The appropriate in vitro maturation time of oocytes was 19-21 h for sheep nuclear transfer,which could significantly improve the quality of blastocysts according to the cell number per blastocyst(P<0.05).     

Effects of in Vitro Maturation Time of Oocytes on Sheep Nuclear Transfer Efficiency

[Objective] The study aimed to provide references for the time of oocyte maturation in vitro and enucleation in the course of sheep nuclear transfer(NT).[Method] Compared the effects of different maturation time of oocytes on enucleation efficiency and reconstructed embryo development by means of blind enucleation and fluorescence microscopy.[Result] Treatment of IVM(in vitro maturation)19-21 h was significantly higher than IVM 16-18 h treatment in oocyte maturation rate(P<0.05)and was significantly higher than IVM 22-24 h treatment in enucleation rate(P<0.05).Three treatments had no significant difference in cleavage rate and blastocyst rate(P>0.05),but IVM 19-21 h treatment was significantly higher than the other 2 treatments in average cell number of blastocysts(P<0.05).[Conclusion] The appropriate in vitro maturation time of oocytes was 19-21 h for sheep nuclear transfer,which could significantly improve the quality of blastocysts according to the cell number per blastocyst(P<0.05).     

卵母细胞体外成熟时间对绵羊核移植效率的影响(英文)

[Objective] The study aimed to provide references for the time of oocyte maturation in vitro and enucleation in the course of sheep nuclear transfer(NT).[Method] Compared the effects of different maturation time of oocytes on enucleation efficiency and reconstructed embryo development by means of blind enucleation and fluorescence microscopy.[Result] Treatment of IVM(in vitro maturation)19-21 h was significantly higher than IVM 16-18 h treatment in oocyte maturation rate(P<0.05)and was significantly higher than IVM 22-24 h treatment in enucleation rate(P<0.05).Three treatments had no significant difference in cleavage rate and blastocyst rate(P>0.05),but IVM 19-21 h treatment was significantly higher than the other 2 treatments in average cell number of blastocysts(P<0.05).[Conclusion] The appropriate in vitro maturation time of oocytes was 19-21 h for sheep nuclear transfer,which could significantly improve the quality of blastocysts according to the cell number per blastocyst(P<0.05).     

Studies on Electrical Activation of Porcine Oocytes Matured in vitro and Embryo Culture Systems

Conditions for electrical parthenogenetic activation of porcine oocytes matured in vitro and in vitro culture systems of porcine embryo were studied. The best results were achieved under the conditions of electrical field strength and the pulse duration at 130Vmm-1/80 μs, with a blastocyst development rate of (20.12 ± 8.18)% (P ＞ 0.05). No significant difference was found between treatments of multiple pulses and a single pulse ( P ＞ 0.05). Parthenogenetic embryos were cultured with different methods and air conditions for 7 days in vitro, blastocyst development rate of embryos with changed culture media [ (26.44 ± 8.35)% ] or changed media with 10% fetal bovine serum (FBS) [ (17.68 ± 5.39)% ] on the fifth day showing no significant difference from that of embryos without change of culture media [ (25.30 ± 7.55) %, P ＞ 0.05 ], while cell numbers of blastocysts from embryos with changed culture media (15.78 ± 5.46 and 14.55 ± 4.81) were significantly lower than number of blastocysts from embryos without change of culture media (18.01 ± 6.79,P ＜ 0.01 ). Blastocyst development rate and blastocyst cell number of embryos cultured in lower O2 (5 % CO2:7%O2:88%N2) also showed no significant difference from those in high O2 (5% CO2 in air) [ (20.78 ± 8.80) % and 17.00 ± 6.12 vs. (25.30 ± 7.55) % and 18.01 ± 6.79, P ＞ 0.05 ]. It is concluded that change of culture media with the same new one or changing over to media with 10% fetal bovine serum (FBS) on the fifth day and low O2 environment are not necessary for porcine embryos development.     

体细胞核移植法获得转人乳铁蛋白基因克隆山羊妊娠的研究(英文)

[Objective] The aim of this study is to understand the effects of donor cell type,embryo stage,number and transfer position on the efficiency of goat transgenic clone.[Method] Using somatic cell nuclear transfer technology,the single goat fetal fibroblasts(GFF)and mammary gland epithelial cells(GMGE)harboring human lactoferrin(hLF)gene were transferred to the enucleated oocyte.Reconstructed karyoplast-cytoplast couplets were fused,activated,and cultured in vitro.Embryos at 2-8 cell stage were transferred into oviduct of synchronized recipients,and blastocysts were transferred into uterine horn.[Result] The pregnancy rate was similar between GFF and GMGE(oviduct transfer:26.47% vs.20.00%),and between oviduct transfer and uterine horn transfer(26.47% vs.25.00%)for GFF group;pregnancy rate in the group with the mean number of embryo transferred per recipient of 21.2 was significantly higher than in those the 5.93 group and 9.64 group(40.00% vs.26.67% and 21.43%).[Conclusion] These results indicate that pregnancy rate of goat transgenic clone couldn't be affected by donor cell type,embryo stage and transfer position but be done by the number of embryo transferred per recipient.In addition,the study also suggests the feasibility of making transgenic goat using GMGE as donor cells.     

Pregnancies Resulting from Transgenic Embryos with Human Lactoferrin Gene Produced by Somatic Cell Nuclear Transfer

[Objective] The aim of this study is to understand the effects of donor cell type,embryo stage,number and transfer position on the efficiency of goat transgenic clone.[Method] Using somatic cell nuclear transfer technology,the single goat fetal fibroblasts(GFF)and mammary gland epithelial cells(GMGE)harboring human lactoferrin(hLF)gene were transferred to the enucleated oocyte.Reconstructed karyoplast-cytoplast couplets were fused,activated,and cultured in vitro.Embryos at 2-8 cell stage were transferred into oviduct of synchronized recipients,and blastocysts were transferred into uterine horn.[Result] The pregnancy rate was similar between GFF and GMGE(oviduct transfer:26.47% vs.20.00%),and between oviduct transfer and uterine horn transfer(26.47% vs.25.00%)for GFF group;pregnancy rate in the group with the mean number of embryo transferred per recipient of 21.2 was significantly higher than in those the 5.93 group and 9.64 group(40.00% vs.26.67% and 21.43%).[Conclusion] These results indicate that pregnancy rate of goat transgenic clone couldn't be affected by donor cell type,embryo stage and transfer position but be done by the number of embryo transferred per recipient.In addition,the study also suggests the feasibility of making transgenic goat using GMGE as donor cells.     

体细胞核移植法获得转人乳铁蛋白基因克隆山羊妊娠的研究(英文)

[Objective] The aim of this study is to understand the effects of donor cell type,embryo stage,number and transfer position on the efficiency of goat transgenic clone.[Method] Using somatic cell nuclear transfer technology,the single goat fetal fibroblasts(GFF)and mammary gland epithelial cells(GMGE)harboring human lactoferrin(hLF)gene were transferred to the enucleated oocyte.Reconstructed karyoplast-cytoplast couplets were fused,activated,and cultured in vitro.Embryos at 2-8 cell stage were transferred into oviduct of synchronized recipients,and blastocysts were transferred into uterine horn.[Result] The pregnancy rate was similar between GFF and GMGE(oviduct transfer:26.47% vs.20.00%),and between oviduct transfer and uterine horn transfer(26.47% vs.25.00%)for GFF group;pregnancy rate in the group with the mean number of embryo transferred per recipient of 21.2 was significantly higher than in those the 5.93 group and 9.64 group(40.00% vs.26.67% and 21.43%).[Conclusion] These results indicate that pregnancy rate of goat transgenic clone couldn’t be affected by donor cell type,embryo stage and transfer position but be done by the number of embryo transferred per recipient.In addition,the study also suggests the feasibility of making transgenic goat using GMGE as donor cells.     

不同条件对绒山羊体外培养初级毛囊生长的影响(英文)

[Objective] The aim of this study is to lay a foundation for illustrating the biological characteristics and growth regulation mechanism of hair follicles.[Method]Cashmere goat primary hair follicles were separated under aseptic condition and cultured in serum-free DMEM and serum-free Williams E media respectively;subsequently,the growth rate and morphological changes were observed under the inverted microscope.[Result]Hair follicles cultured in serum-free DMEM media showed a growth rate of 0.034 mm/d during the first 3 days,whose structure and morphological characteristics could maintian a stable status for a long time in the growth process.Hair follicles grew much faster in the serum-free Williams E media with a growth rate of 0.077 mm/d during the first 3 days.[Conclusion]There were significant differences(P<0.05)between the growth of cashmere goat hair follicles cultured in the 2 kinds of media.Serum-free Williams E medium was superior to serum-free DMEM medium.     

Effect of GPAG Supplement on Porcine Oocyte Maturation and Its Following Embryonic Development by Parthenogenetic Activated

The aim of this study was to compare the effect of GPAG and commonly used FCS on porcine oocyte maturation and subsequent embryonic development after the fertilization. COCs were aspirated from follicles and cultured for 16, 24, 32, 40 and 48 h in TCM-199 medium either with GPAG or FCS. After 24 h with GPAG, 89.4% of oocytes reached M Ⅰ stage while in the medium supplemented with FCS, only 27.7% of oocytes reached the same stage （P〈0.05）. Prolonged incubation for up to 32 h clearly demonstrated that some of oocytes cultured in GPAG medium were at M Ⅱ stage （35.7%）, few of oocytes from FCS medium were at M Ⅱ stage （7.5%） （P〈0.05）. Both groups of oocytes reached the same stage of maturation within 48 h. After 48 h of culture, the oocytes with extruded polar bodies were inseminated. Fertilized oocytes were cultured in PZM3 medium supplemented with 3 mg.mL of BSA. After 7 days, the development and the quality of embryos were evaluated. The results showed that the maturation of oocytes in the presence of GPAG significantly increased their subsequent developmental ability when compared with FCS supplementation （29.2% ： 18.9% of blastocysts, P〈0.05）. However, differential staining revealed that once blastocysts were formed in either group, they had the same total cell number （39 ： 38） and the ICM/total cell ratio （0.26 ： 0.28）     

中国农业科学

In this experiment, the bovine follicular oocytes were aspirated from the ovaries of Chinese Holsteins with laparoscope made in China. The results were as following: for identifying the suitable negative aspiration pressure, six different pressures (50, 100, 150, 200, 250 and 300mmHg)were tested. The aspiration pressure of 100mmHg was the best. Its oocyte recovery rate was 37. 2%, and G I , G Ⅱ oocyte rate was 89. 5%. The heifers were picked up by laparoscope once or twice a week. Each heifer was collected with 2. 4 oocytes once a week or 4. 4 oocytes twice a week.Its oocyte recovery rate was 48. 0% and the G Ⅰ ,G Ⅱ oocyte rate was 93. 5%. In addition, 1.9 oocytes were collected from each cow once a week or 5.4 oocytes from each cow twice a week. Its oocyte recovery rate was 51.7% and the G Ⅰ , G Ⅱ oocyte rate was 85. 1%. It showed that it was possible to pick up bovine oocyte twice a week. Two cows were picked up twice a week for several weeks(53 times). 268 follicles were aspirated(5.1 follicles per cow per time), and 141 oocytes were recovered(2.7 oocytes per cow per time). The oocyte recovery rate was 52.5%, and the G Ⅰ , G Ⅱ oocyte rate was 85. 1%. It was advisable to pick up oocytes twice a week continuously. Some cows in estrous cycles were superovulated with PMSG(500IU). Each of them could be recovered 2.3 follicles and 1.1 oocytes, the others were superovulated with FSH(0. 7mg) , each of them could be aspirated with 4.4 follicles and 2.3 oocytes. It was obvious that the effect of OPU(oocyte pick up) by superovulation with FSH was much better than that with PMSG. The best time for OPU with laparoscope was at the beginning of cow's estrous cycles. At the first day of their estrus, each of them could be averagely aspirated with 8 follicles and 5.7 oocytes.     

“胰岛素-转铁蛋白-硒钠”对山羊卵母细胞体外成熟及胚胎发育的影响

[目的]为提高山羊卵母细胞及胚胎体外培养的效率,探索无血清体外培养体系。[方法]在目前卵母细胞体外成熟及胚胎培养所用的常规培养液中添加1%ITS(胰岛素-转铁蛋白-硒钠)或以1%ITS分别替代2种培养液中FBS,对山羊卵母细胞及孤雌激活胚胎进行体外培养,检测ITS对其发育率的影响。[结果]添加ITS到卵母细胞成熟液中,对卵母细胞成熟率没有明显提高,但显著提高了其激活后孤雌胚胎的囊胚率(58.06%vs.48.19%);以ITS替代成熟液中FBS,取得了与FBS组相似的成熟率、卵裂率和囊胚率。添加ITS到胚胎培养液中,显著提高了山羊孤雌胚胎的囊胚率(68.30%vs.56.82%);以ITS替代胚胎培养液中FBS,卵裂率与FBS组无显著差异,囊胚率显著低于FBS组(42.33%vs.56.82%)。[结论]ITS对山羊卵母细胞体外成熟及胚胎早期发育均有促进作用;另外,ITS可以替代卵母细胞成熟培养液中的血清,作为无血清培养体系用于相关研究。     

尿嘧啶和ATP对牛卵母细胞成熟及激活后孤雌胚胎发育的影响

[目的]探究尿嘧啶(Uracil)和三磷酸腺苷二钠盐(ATP)对牛卵母细胞成熟及激活后孤雌胚胎发育的影响。[方法]在体外成熟培养液中添加不同浓度的尿嘧啶和ATP,研究其对牛卵母细胞体外成熟及激活后孤雌胚胎发育能力的影响。[结果]在成熟培养液中添加50μg/ml尿嘧啶能够显著提高卵母细胞的成熟率、分裂率及孤雌胚胎囊胚率;在成熟培养液中分别添加0、250、500、750μg/mlATP,对卵母细胞成熟率影响不大,但添加500μg/ml ATP能显著提高激活后孤雌胚胎囊胚率。[结论]该研究为牛卵母细胞IVM研究提供了参考资料。     

CB·氨基酸在猪卵母细胞孤雌激活及体外培养中的作用

[目的]研究CB、氨基酸在猪卵母细胞孤雌激活及体外培养体系中的作用,为优化相关技术体系提供依据。[方法]卵母细胞经体外成熟培养,采用不同孤雌激活方法(Ele.+CB组、CB组、Ele.组、对照组),研究CB在激活中的作用。激活后采用PZM3培养液,并去除氨基酸成分,探索氨基酸对体外培养胚胎的作用。[结果]CB+Ele.组的卵裂率显著地高于其他3组(P<0.05),囊胚率为32.7%,囊胚细胞数为61.4,而其他3组均未出现囊胚。电激活(脉冲电压100 V/mm,脉冲时程100μs,脉冲次数1次)联合CB处理,可激活卵母细胞体外发育至囊胚,CB有助于提高卵裂率、囊胚率。添加氨基酸的培养液中的卵母细胞的卵裂率、囊胚率、囊胚细胞数显著提高(P<0.05),表明氨基酸能提高猪孤雌激活胚胎培养效果。[结论]添加CB、氨基酸在猪卵母细胞孤雌激活及体外培养体系中能提高胚胎的培养效果。     

不同浓度Leptin对猪卵母细胞体外成熟和早期孤雌发育的影响

[目的]系统比较了在成熟培养液中添加Leptin对猪卵母细胞体外成熟和孤雌激活胚的发育率和囊胚质量,同时比较了成熟培养液和胚胎培养液添加Leptin对孤雌激活胚发育的影响。[方法]成熟培养液中添加不同浓度Leptin对猪卵母细胞体外成熟和早期孤雌发育能力的影响及囊胚的内细胞团细胞数目,并且研究了成熟培养液和胚胎培养液单独或联合添加Leptin对猪早期孤雌胚胎发育能力的影响及囊胚的内细胞团细胞数目。[结果]成熟培养液中添加不同浓度Leptin(0,10,50,100和200 ng/ml),体外成熟44 h,卵母细胞核成熟率统计分析差异不显著(P>0.05);将核成熟卵母细胞进行化学激活,第2天胚胎卵裂率统计分析差异不显著(P>0.05),但100ng/ml组与其他组第7天的囊胚率及囊胚的总细胞数差异显著(P<0.05);成熟液添加Leptin 100 ng/ml、胚胎培养液添加Leptin 10 ng/ml与其他组第7天的囊胚率及囊胚的内细胞团细胞数差异显著(P<0.05)。[结论]Leptin对卵母细胞成熟和孤雌胚胎发育能力上有相辅相成的作用。     

血管内皮生长因子对水牛卵母细胞体外受精的影响

[目的]探讨血管内皮生长因子（Vascular endothelial growth factor,VEGF）对水牛卵母细胞体外受精的影响,为优化水牛卵母细胞体外培养体系及提高水牛体外胚胎生产效率奠定基础.[方法]以带有3层以上卵丘细胞的卵丘—卵母细胞复合体（COCs）为材料,分别在水牛卵母细胞体外成熟液和受精液中添加重组人VEGF165,研究VEGF对水牛卵母细胞体外成熟、体外受精和早期胚胎发育的影响.[结果]在成熟液中添加10ng/mLVEGF,水牛卵母细胞的第一极体排出率由57.3％提高到72.3％,差异显著（P＜0.05,下同）;体外受精后的受精卵分裂率由41.0％显著提高到53.3％.在受精液中添加10 ng/mL VEGF,水牛体外受精的受精卵分裂率由41.9％提高到63.0％,差异极显著（P＜0.01,下同）,囊胚形成率由17.0％显著提高到32.6％.在成熟液和受精液中同时添加10ng/mLVEGF,水牛体外受精的受精卵分裂率和囊胚形成率由43.4％和19.0％极显著提高到64.3％和27.2％,囊胚细胞数由89.5显著提高到106.8.[结论]VEGF能促进水牛卵母细胞体外成熟、体外受精及早期胚胎的发育,因此可将VEGF用于水牛卵母细胞体外受精体系的优化,提高水牛体外胚胎生产效率.     

培养液对猪卵母细胞体外成熟及孤雌胚胎早期体外发育的影响

[目的]提高卵母细胞体外成熟质量,优化猪早期胚胎体外培养体系。[方法]以改良TCM199,NCSU-23培养液为基础液,分别添加10%的猪卵泡液（PFF）和10%的优质胎牛血清（FBS）进行卵母细胞体外成熟培养,以成熟率、孤雌胚胎体外发育率等为指标,研究不同培养液对猪卵母细胞体外成熟及孤雌胚胎体外发育的影响。[结果]猪卵母细胞在TCM199,TCM199＋FBS和TCM199＋PFF组成熟42 h的成熟率分别为（54.2±3.5）%、（68.5±3.2）%和（69.3±3.7）%,在NCSU-23,NCSU-23＋FBS和NCSU-23＋PFF组成熟42 h的成熟率分别为（51.6±3.3）%、（63.2±3.1）%和（65.5±3.5）%,添加10%的FBS和PFF在0.05水平上显著提高了卵母细胞成熟率;6组不同成熟培养液得到的成熟卵母细胞在孤雌激活后囊胚发育率差异不显著,但TCM199＋PFF组的囊胚细胞数（36.5±4.8）在0.05水平上显著高于TCM199和NCSU-23组的囊胚细胞数（18.7±3.2和15.5±2.4）。[结论]改良TCM199,NCSV-23培养液中添加10%PFF和10%FBS可显著促进猪卵母细胞体外成熟及孤雌胚胎早期体外发育。     

卵泡液对牛卵母细胞发育潜力的影响

为了确定卵泡液(直径大于10mm)对卵母细胞成熟的影响,共收集1759枚卵母细胞进行试验。在成熟液中分别添加0%,10%,30%和50%的卵泡液(bFF,bovinefollicularfluid),成熟培养24h后,孤雌激活或者体外受精和胚胎培养;对比10?F和10%胎牛血清(FBS),以及观察微滴培养对卵母细胞成熟的影响。结果表明,在孤雌激活组中,bFF浓度对卵裂率的影响差异不显著,但胚胎囊胚发育率高于对照组(P<0.05);在体外受精组中随着bFF浓度增加而卵裂率下降(P<0.01),而且囊胚率也下降(P<0.05)。在10%的bFF组中,孤雌激活囊胚率和体外受精囊胚率分别为47.1%和38.0%,囊胚扩张率显著高于其它组。从试验得出,添加10?F有利于卵母细胞胞质成熟,且不会降低到受精率,而且可以替代FBS;微滴培养不利于卵母细胞成熟。     

乙二胺四乙酸钠、能量基质、氨基酸对猪卵母细胞体外成熟和早期孤雌发育的影响

猪的卵巢卵母细胞在含不同剂量葡萄糖+丙酮酸钠的成熟培养基中体外成熟培养44-48 h,检查核成熟率(试验一),进行化学法孤雌激活后,在含不同浓度的乙二胺四乙酸钠(EDTA-Na)和不同剂量的葡萄糖+谷氨酰胺+牛磺酸+亚牛磺酸的培养基中进行体外培养,检查孤雌激活胚第48小时的卵裂率和第168小时的囊胚发育率(试验二),研究成熟培养基中葡萄糖+丙酮酸钠剂量对卵母细胞体外成熟核成熟率的影响及培养基中不同浓度EDTA-Na和葡萄糖+谷氨酰胺+牛磺酸+亚牛磺酸的剂量对孤雌激活胚胎体外发育的影响.结果表明:(1)葡萄糖+丙酮酸钠在1倍剂量时,猪卵母细胞体外成熟的核成熟率为(59.3±5.0)%;当剂量加倍时,核成熟率极显著下降(P<0.01),为(51.0±4.4)%.(2)当EDTA-Na浓度增高,囊胚发育率上升,当浓度达50μmol.L-1时囊胚发育率最高,为(7.2±4.1)%,此后随浓度升高囊胚发育率下降;添加适当浓度(25-100μmol.L-1)EDTA-Na对猪孤雌激活胚的囊胚发育有利(P<0.05),当浓度达200μmol.L-1时其有利作用消失.(3)培养基中葡萄糖+谷氨酰胺+牛磺酸+亚牛磺酸的剂量过高对卵裂率无显著影响(P>0.05),但对囊胚发育不利(P<0.05).可见:(1)适当浓度(25-100μmol.L-1)EDTA-Na对猪早期胚胎体外发育具有促进作用;(2)成熟培养基中能量基质剂量过高会抑制猪卵母细胞体外成熟;(3)胚胎培养基中葡萄糖+谷氨酰胺+牛磺酸+亚牛磺酸的剂量过高会抑制猪早期胚胎的体外发育.     

亮甲酚蓝染色对水牛卵母细胞体外受精和孤雌激活胚胎发育的影响

[目的]探讨亮甲酚蓝（BCB）染色对水牛卵母细胞体外受精及孤雌激活胚胎发育的影响。[方法]卵丘卵母细胞复合体（COCs）在体外成熟培养前用浓度26μmol/L的亮甲酚蓝染色90 min,然后根据卵母细胞胞质蓝色的有无分为阳性（BCB＋,蓝色）和阴性组（BCB-,不着色）。以未经染色处理的COCs为对照组,控制对照组卵母细胞在含0.4%牛血清白蛋白（BSA）的杜氏磷酸盐缓冲液（DPBS）中孵育90 min。体外成熟培养后统计各组的成熟率、体外受精和孤雌激活后的发育率。[结果]BCB＋组的成熟率（65.70%）显著高于对照组（59.86%）、控制对照组（58.42%）和BCB-组（48.86%）。在体外受精后的囊胚发育率方面,BCB＋组（27.08%）和对照组（25.49%）差异不显著,但显著高于控制对照组（20.50%）和BCB-组（8.45%）。在孤雌激活后的囊胚发育率方面,BCB-组显著低于其他各组,但BCB＋组、对照组以及控制对照组之间无显著差异。[结论]亮甲酚蓝染色筛选出的阳性卵母细胞具有较好的核成熟发育潜能,但并不能显著提高卵母细胞体外受精及孤雌激活后的胚胎发育率。     

微滴中胚胎数量对猪孤雌胚早期体外发育的影响

[目的]优化猪早期胚胎培养体系,为单精子显微受精及体细胞核移植等研究提供依据。[方法]以猪的早期孤雌胚胎为材料,分别将人工激活后的70、50、30、15和5枚卵母细胞放入100μlNCSU-23培养液的微滴中进行培养,探讨了胚胎培养数量对猪早期孤雌胚发育的影响。[结果]100μl的培养微滴中培养30、50和70胚胎组,其分裂率分别为64.0%、65.2%和67.1%,显著高于5胚胎组,而与15胚胎组的差异不显著 在囊胚率方面,70胚胎组的最高,为3.0%,与50胚胎组的（1.7%）无显著差异,与5、15和30胚胎组的差异显著 各组的囊胚细胞数之间没有显著差异。[结论]在该研究条件下,当微滴体积为100μl时,培养50~70枚猪孤雌胚胎的效果最好,即,胚胎数∶培养滴体积=（1∶1.43~2.00）。