兔原核胚体外序贯培养

采用添加 10 % FBS的 RPMI16 4 0培养液和 m RPMI16 4 0培养液对兔原核期受精卵进行了体外序贯培养 ,并与添加 10 % FBS的 RD培养液单一培养作了比较。结果显示 :体外培养至 72 h时 ,2个培养组 8-细胞胚率、桑葚胚率和囊胚发育率无显著差异 (P>0 .0 5 ) ,但 RD单一培养组已有 10 .3%出现退化 ,与序贯培养组之间差异显著 (10 .3%比0 ,P<0 .0 5 ) ;体外培养 16 8h,序贯培养组和 RD培养组的贴附率分别为 5 9.7%和 4 1.4 % ,外延生长率分别为 4 3.1%和 2 2 .4 % ,桑葚胚率为 10 0 %和 89.7% ,脱带率为 33.3%和 5 .2 % ,二者之间贴附率、外延生长率差异显著 (P<0 .0 5 ) ,桑葚胚率、脱带率差异极显著 (P<0 .0 1) ;序贯培养 96 h的囊胚细胞数为 (12 4 .6± 6 .36 )个 /枚 ,RD培养同期囊胚细胞数为 (118.2± 5 .2 5 )个 /枚 ,差异显著 (P<0 .0 5 )。结果表明 ,序贯培养能够有效克服兔早期胚胎发育阻滞 ,促进胚胎的正常生长发育 ,提高胚胎质量 ,并能促进胚胎的孵化和附植     

热应激对植入前牛胚胎发育的影响

采集屠宰母牛卵巢和输卵管中的卵母细胞，进行体外受精，受精卵继续在体外培养。将发育3d的牛胚胎于42℃下分别培养0．5、2．0和4．0h，与对照组（39℃）牛胚胎相比，接触最高热应激（42℃，4h）处理的胚胎，约509／6的胚胎发育速度明显下降。而处理组牛胚胎发育到第9d的囊胚数、细胞数及内细胞团与滋养外胚层比率与对照组差异不显著（P〉0．05）。表明，温度升高对牛囊胚期胚胎的发育没有明显损害。对9d的囊胚进行基因性别分析发现，正常体外培养的雄胚发育快于雌胚，而在发育第3d受42℃热应激后，到囊胚期生存的胚胎性别比率发生改变，雌胚比率高于雄胚，表明雌性胚胎比雄性胚胎具有更强的抗热应激能力。     

2细胞期胚胎采集时间对电融合方法体外制备牛四倍体胚胎效率的影响

通过检测牛早期胚胎分裂速率的时间分布,并将体外受精后0~34 h与26~30 h时间段内采集的2细胞期胚胎分别作为电融合对照组与电融合试验组。将对照组与试验组的2细胞期胚胎分别用于电融合,并将电融合后的胚胎继续体外培养至囊胚期,检测并对比分析2组胚胎的融合率、分裂率、囊胚率和四倍体制备率。结果显示,牛2细胞期胚胎在体外受精后28~30 h其内分裂速率达到最高峰(18.9%)。电融合对照组的融合率、分裂率略高于试验组胚胎(85.2%vs.84.2%,82.2%vs.80.9%),试验组的囊胚率略高于电融合对照组(45.4%vs.44.8%),但差异均不显著。而电融合试验组的四倍体制备率显著高于电融合对照组(28.3%vs.14.5%)。结果表明,牛2细胞期胚胎的采集时间对电融合方法体外制备牛四倍体胚胎的效率有显著影响。     

小鼠胚胎DNA氧化损伤模型的建立

为了研究胚胎DNA损伤修复机制,试验以H2O2为DNA损伤诱导剂,建立小鼠胚胎DNA氧化损伤模型,采用不同浓度H2O2培养小鼠受精卵,观察其对小鼠胚胎发育能力的影响;再通过免疫荧光方法检测γH2AX和ρATM蛋白的表达状况,以判断H2O2处理所导致的胚胎DNA损伤情况及较适宜的诱导条件。结果表明:1.5~2.4μmol/L H2O2处理对小鼠胚胎发育能力具有影响,与KOSM(-)对照组相比,H2O2处理组小鼠胚胎的分裂率、囊胚率呈下降趋势,其中1.8~2.4μmol/L H2O2处理组的胚胎分裂率、囊胚率呈显著下降趋势(P0.05)。免疫荧光法检测发现KOSM(-)对照组中,2-细胞期胚胎的γH2AX焦点数多于1-细胞期胚胎,ρATM在两个时期的胚胎中未检测到。γH2AX和ρATM在1.5,1.8μmol/L H2O2处理组的1-细胞期胚胎中均未检测到;2-细胞期胚胎表达γH2AX,未检测到ρATM。2.1,2.4μmol/L H2O2处理组的1-细胞和2-细胞期胚胎均有γH2AX和ρATM表达,其中2.4μmol/L H2O2处理组两个时期胚胎的γH2AX焦点数多于2.1μmol/L H2O2处理组。说明1.5~2.4μmol/L H2O2处理会影响小鼠胚胎体外发育能力,2.1,2.4μmol/L H2O2处理可诱导小鼠胚胎发生DNA双链断裂现象。     

早期胚胎体外培养微流控芯片的制备及初步应用

利用微流控芯片模拟输卵管微环境,探讨物理性刺激对早期胚胎发育的影响,为解决早期胚胎体外囊胚发育率较低、胚胎质量差的问题提供帮助。本实验采用一次铸造成型制备微流控芯片培养装置,并应用培养装置对小鼠1-细胞期胚胎进行培养,观察胚胎体外发育率。结果表明,利用微流控芯片培养组与对照组2相比,2-细胞胚胎发育率差异不显著(P0.05),但是,在8-细胞胚胎发育率、桑葚胚发育率和囊胚发育率都差异显著(P0.05)。此外,囊胚内总细胞数显著提高(P0.05)。结论:一次铸造成型微流控芯片制作方法简便易行且培养液不易外漏,这种培养装置能显著提高小鼠早期胚胎体外囊胚发育率和胚胎的质量。     

β-巯基乙醇或牛磺酸对牛体外受精后早期胚胎的影响

对屠宰黄牛的卵母细胞经体外成熟(IVM)、体外受精(IVF)后的早期胚胎,在β-巯基乙醇(-βME)或牛磺酸等添加物的胚胎培养液中的后续发育进行了研究,并探讨了其影响因素,以期筛选出最佳的体外培养条件。试验结果表明:4~8细胞期添加-βME可显著提高胚胎桑椹胚、囊胚发育率和囊胚细胞数,但不能改善孵化囊胚的质量。在体外发育培养液中添加7mM牛磺酸可显著提高桑椹胚率和囊胚率(P<0.05),并且在4~8细胞期添加牛磺酸最为合适。     

5-Aza-CdR对德保黑猪手工克隆重构胚胎体外发育效果的影响

为了探讨5-氮杂-2'-脱氧胞苷（5-Aza-CdR）对德保黑猪手工克隆（HMC）重构胚胎体外发育效果的影响，本研究分别从供体细胞和重构胚入手，比较了5个不同处理浓度（0、5、10、20和40 nmol/L）5-Aza-CdR处理HMC重构胚的体外发育效果，筛选最佳处理浓度；在最佳浓度下比较5个不同处理时间（0、24、48、72和96 h）对HMC重构胚的体外发育效果，筛选最佳处理时间；用4个不同浓度（0、0.25、0.5和1 μmol/L）5-Aza-CdR结合最佳浓度和最佳时间处理供体和重构胚，比较其体外发育潜能。结果显示，与空白对照组相比，5、10、20和40 nmol/L 5-Aza-CdR处理72 h对重构胚卵裂率均无显著差异（P>0.05），20 nmol/L 5-Aza-CdR处理能显著提高重构胚的囊胚率（P<0.05），10和20 nmol/L 5-Aza-CdR处理均能显著提高囊胚细胞数（P<0.05），其中以20 nmol/L 5-Aza-CdR效果最佳；与空白对照组相比，利用20 nmol/L 5-Aza-CdR处理HMC重构胚72 h能显著提高重构胚的囊胚率和囊胚细胞数（P<0.05），其余处理时间对重构胚卵裂率、囊胚率和囊胚细胞数均无显著影响（P>0.05）；在囊胚的最佳处理浓度（20 nmol/L）和最佳处理时间（72 h）下，结合供体的4个处理浓度（0、0.25、0.5和1 μmol/L），同时处理重构胚和供体，各处理组HMC重构胚的发育潜能均有提高，但效果均不显著（P>0.05），其中0.25~0.5 μmol/L 5-Aza-CdR处理效果较佳。综上表明，适宜浓度（0.25~0.5 μmol/L）的DNA甲基化酶抑制剂5-Aza-CdR处理供体细胞72 h并结合20 nmol/L 5-Aza-CdR处理重构胚72 h均能有效提高德保黑猪HMC重构胚胎的体外发育潜能，该结果可为今后研究德保黑猪HMC胚胎DNA甲基化调控机制提供参考。     

曲古抑菌素A处理克隆胚对囊胚发育率的影响

[目的]探讨曲古抑菌素A（Trichostatin A,TSA）处理对囊胚发育率的影响。[方法]以牛胎儿成纤维细胞作为供体核,以牛卵母细胞作为受体胞质进行体细胞核移植,用80 nmol/L TSA处理供体细胞12 h,核移植后继续处理克隆胚胎0、6、12和24 h,应用激光共聚焦显微镜检测供体细胞组蛋白H4K12乙酰化水平,并通过核移植检测TSA不同时间处理的克隆胚胎囊胚发育率。[结果]TSA处理12 h的供体细胞组蛋白H4K12乙酰化水平显著增高（P〈0.05）;80 nmol/LTSA处理12 h的克隆胚的囊胚发育率（21.9%）高于未处理组（16.5%）,差异显著（P〈0.05）。[结论]供体细胞和克隆胚胎经TSA处理的12 h的克隆胚胎,显著提高了体外发育能力。     

SAHA处理对猪手工克隆胚胎体外发育潜能的影响

为了探讨组蛋白去乙酰化酶抑制剂辛二酰苯胺异羟肟酸(SAHA)对德保猪手工克隆胚胎(HMC)发育潜能的影响,试验摸索SAHA的适宜处理浓度[0(对照),1.0,2.5,5.0,7.5,10.0μmol/L]和时间(0,6,12,24 h);之后分为4组,SAHA组、体外受精(IVF)组、孤雌激活(PA)组、对照(HMCC)组,分别在体外发育的1细胞期、2细胞期、4细胞期、囊胚期收集胚胎,在相同时期下比较各组胚胎组蛋白H4K8乙酰化(Ac H4K8)水平差异和相关基因(HDAC1、HAT1、ASF1A、OCT-4)相对表达量。结果表明:7.5μmol/L SAHA处理囊胚率显著高于对照(P0.05),12 h囊胚率显著高于0 h(P0.05);所以适宜处理浓度为7.5μmol/L,适宜处理时间为12 h。在1细胞期、2细胞期、囊胚期,SAHA组Ac H4K8水平接近IVF组水平(P0.05)。在囊胚期,SAHA组HDAC1基因相对表达量接近IVF组(P0.05);在囊胚期,SAHA组OCT-4基因相对表达量接近IVF组(P0.05)。说明SAHA可以使HMC胚胎Ac H4K8水平接近IVF水平,并纠正克隆胚胎乙酰化的异常,从而提高克隆胚胎发育潜能。     

Scriptaid对猪移植体细胞核在胚胎外发育的影响

组蛋白去乙酰化酶(histone deacetylases,HDACs)是调控基因的关键蛋白酶,乙酰化是一种可逆的蛋白共价修饰。组蛋白去乙酰化酶可以改变特定基因的转录和表达水平,诱导细胞的分化和凋亡。为了检测一种低毒新型的组蛋白去乙酰化酶抑制剂Scriptaid对克隆猪胚胎发育的影响,进行了不同浓度和不同时间的处理,并在体外检测了胚胎分裂率和囊胚发育能力。研究以对照试验作为基础,以凌源禾丰种猪场长白猪胎儿成纤维作为供体细胞,然后通过体细胞核移植(SCNT)技术获得体外发育胚胎。将重构胚胎以不同浓度(0、200、500、700、900 nmol/L)Scriptaid处理后,并进行不同时间(0、15、36、72 h)的培养,然后通过观察不同处理浓度和处理时间下胚胎在2细胞阶段分裂率和囊胚发育率。再通过Real-time PCR检测2细胞阶段未处理组与Scriptaid处理组的Oct4、Sox2两个因子的相对表达变化。通过统计学分析发现,与未处理组相比,500 nmol/L Scriptaid处理15 h囊胚发育率显著增加(P0.05),但2细胞阶段分裂率基本不变,而未处理组克隆胚胎在囊胚期Oct4基因表达的倍数明显低于经过Scriptaid处理的克隆胚胎在囊胚期的表达倍数(P0.05),这说明经过500 nmol/L Scriptaid处理后,克隆胚胎的Oct4表达量明显提高。未经过处理的克隆胚胎在囊胚期Sox2基因的表达倍数和经过Scriptaid处理的克隆胚胎在囊胚期Sox2基因的表达倍数有差异但是差异不明显,这说明经过500 nmol/L Scriptaid处理之后,基因Sox2表达虽然有提高,但总体来说和未处理的胚胎相比差异不大(P0.05)。     

The Effects of Blastomere Biopsy and Oxygen Tension on Bovine Embryo Development, Rate of Apoptosis and Interferon-τ Secretion

A series of experiments was performed to examine the effects of blastomere biopsies on subsequent development of IVF-derived bovine embryos. The first experiment was designed to assess the optimal time for blastomere removal. One blastomere was removed either 48 or 72 h after IVF. Biopsy at 48 h resulted in 17.2% of embryos proceeding to the blastocyst stage, which was lower than when biopsies were performed at 72 h (37.5%, p < 0.05). In the second experiment, embryos were cultured either under atmospheric or 5% O(2) following blastomere removal. Biopsies had no effect on rate of blastocyst formation with 36% of controls and 33.7% of biopsied embryos proceeding to that stage. However, culture under 5% O(2) significantly increased the number of blastocysts from 29.9% to 40.3% (p < 0.05). This effect was significant in both biopsied and control embryos. In the final experiment, biopsied embryos were again cultured under different oxygen tension. Blastocysts were collected and cultured individually for 48 h in medium droplets in their respective O(2) concentration after which time the medium was assayed for concentration of interferon-tau (IFN-tau). Reduced O(2) concentration again significantly increased blastocyst formation from 24.9% to 41.9% (p < 0.05). IFN-tau secretion was not affected by biopsies, but culture under atmospheric O(2) resulted in significantly increased IFN-tau concentration in medium droplets (12274.0 +/- 2825.9 pM vs 5046.5 +/- 2562.2 pM; p < 0.05).     

Oviduct epithelial cell co-culture of early porcine embryos.

One- to 16-cell porcine embryos were cultured in either Whittens medium supplemented with bovine serum albumin and fetal calf serum (WM) or in the same medium with porcine oviduct epithelial cell co-culture (WM-Poec). All stages of embryos cultured in WM-POEC had higher cell counts after 144-168 h of development than did embryos in WM. There was however, no significant difference in blastocyst formation rate of embryos cultured in WM-POEC over those cultured in WM. A high proportion of the embryos entering culture at the 1-2-cell were able to pass the 4-cell block stage in both WM and WM-POEC, 81% and 77%, respectively. In both media, most of the 1-2-cell embryos arrested their development at the compacted morula stage and failed to blastulate while embryos initiating culture at the 4- and 8-16-cell embryos formed blastocysts in culture at a rate of 80-90%.     

OPS玻璃化冷冻对小鼠四倍体胚胎体外发育的影响

为探究开放式拉长细管(OPS)玻璃化冷冻对四倍体胚胎发育的影响,本实验利用2-细胞胚胎电融合法制备四倍体胚胎,再对四倍体胚胎进行OPS玻璃化冷冻,分别观察记录二倍体胚胎、四倍体胚胎以及冷冻解冻后四倍体胚胎的发育情况。结果表明:2-细胞胚胎电融合效率为96.1%;二倍体胚胎组与电融合后四倍体胚胎组的囊胚率和孵化囊胚率差异不显著;冷冻解冻后四倍体胚胎的囊胚率(100%)与四倍体新鲜组(93.3%)差异不显著,其孵化囊胚率(72.3%)较新鲜组(64.9%)显著增高(P<0.05);四倍体冷冻解冻组的囊胚细胞数(31.96)与新鲜组(32.54)无显著差异;冷冻解冻后的四倍体早期囊胚进行体外培养时其发育速度比对照组更快。可见,冷冻对小鼠四倍体胚胎的囊胚率和囊胚细胞数均无显著影响,但孵化囊胚率显著提高,且OPS玻璃化冷冻后使四倍体胚胎的发育速度更快。     

Effects of BSA and fetal bovine serum in culture medium on development of rat embryos

Rat 1-cell embryos, recovered from naturally mated females, were cultured in a chemically defined medium (mR1ECM) or in mR1ECM supplemented with BSA (4 mg/ml; mR1ECM-BSA) or fetal bovine serum (FBS; 10%, v:v; mR1ECM-FBS) instead of polyvinylalcohol. There was no difference in percentages of embryos that developed to the 2-cell to blastocyst stages between mR1ECM and mR1ECM-BSA, but in mR1ECM-FBS, no development beyond the 2-cell stage was observed. When embryos were transferred to mR1ECM-FBS from mR1ECM after 24 to 64 h of culture, development of embryos to and beyond the 4-cell stage was inhibited. However, when transferred after 80 h of culture, more embryos developed to blastocysts and hatching or hatched blastocysts than in embryos cultured in mR1ECM. When 8-cell embryos and early morulae obtained after 72 and 80 h of culture in mR1ECM, respectively, were cultured in mR1ECM-FBS, a higher proportion of early morulae developed to the blastocyst stage than did 8-cell embryos. When morulae obtained after culture in mR1ECM or mR1ECM-BSA were transferred to recipient females, there was no difference in proportions of fetuses obtained. However, a higher proportion of blastocysts cultured in mR1ECM-FBS developed to fetuses compared with those obtained in mR1ECM. These results indicate that BSA has neither deleterious nor beneficial effects on development of rat 1-cell embryos. In contrast, FBS has deleterious effects on early cleavage of embryos but it promotes more rapid development of morulae to blastocysts, resulting in better quality blastocysts.     

猪孤雌激活早期胚胎线粒体分布及mtDNA拷贝数变化的研究

本研究通过线粒体分子探针标记技术检测孤雌激活早期胚胎线粒体的分布变化,运用实时荧光定量PCR技术检测mtDNA拷贝数的变化,揭示早期胚胎发育过程中线粒体分布、mtDNA拷贝数变化趋势。结果表明,成熟卵母细胞电激活后,由2-细胞胚胎开始,卵裂球内线粒体分布均匀且密集,每个细胞均有分布,卵裂球之外的空隙未见线粒体分布,直到囊胚形成,线粒体均有分布。孤雌激活4-细胞胚胎mtDNA拷贝数显著高于8-细胞胚胎mtDNA拷贝数(907210.77±145520.77,186224.33±103308.00,P<0.05),但显著低于2-细胞胚胎、桑椹胚、囊胚的mtDNA拷贝数(1563422.54±224666.51、1697626.25±176999.53和1752301.29±101146.64,P<0.05)。孤雌激活扩张囊胚mtDNA拷贝数最高,为2812545.67±156819.31,显著高于其他发育时期胚胎的mtDNA拷贝数(P<0.05)。由此可见,孤雌激活早期胚胎发育进程中线粒体分布及mtDNA拷贝数会发生变化。     

Epidermal growth factor improves developmental competence and embryonic quality of singly cultured domestic cat embryos

This study examined the influence of EGF on the expression of EGF receptors (EGFR) and developmental competence of embryos cultured individually versus those cultured in groups. Cat oocytes were in vitro matured and fertilized (IVM/IVF), and cleaved embryos were randomly assigned to one of seven culture conditions: one group each in which embryos were subjected to group culture supplemented with or without 5 ng/ml EGF and five groups in which embryos were subjected to single-embryo culture supplemented with EGF (0, 5, 25, 50 or 100 ng/ml). Morulae, blastocysts and hatching blastocysts were assessed at days 5 and 7; post IVF, respectively, and total blastocyst cell numbers were assessed at day 7. Relative mRNA expressions of EGFR of 2–4-cell embryos, 8–16-cell embryos, morulae and blastocysts cultured in groups or singly with or without EGF supplementation were examined. OCT3/4 and Ki67 in blastocysts derived from the group or single-embryo culture systems with or without EGF supplementation were localized. A higher rate of embryos cultured in groups developed to blastocysts than individually incubated cohorts. Although EGF increased blastocyst formation in the single-embryo culture system, EGF did not affect embryo development in group culture. Expression levels of EGFR decreased in morulae and blastocysts cultured with EGF. An increased ratio of Ki67-positive cells to the total number of cells in the blastocyst was observed in singly cultured embryos in the presence of EGF. However, EGF did not affect the expression of OCT3/4. These findings indicate that EGF enhanced developmental competence of cat embryos cultured singly by stimulating cell proliferation and modulating the EGFR expression at various developmental stages.     

The blastocyst production rate and incidence of chromosomal abnormalities by developmental stage in in vitro produced porcine embryos

The present study was conducted to determine the relationship between embryonic development speed at different stages (the cleaved stage at 52 h and the blastocyst stage at 6 days post insemination) and incidences of chromosome abnormalities in in vitro produced porcine embryos. Porcine oocytes were collected from 3-6-mm ovarian follicles obtained at a slaughterhouse and matured in modified NCSU-37 medium for 44-46 h. Following in vitro fertilization with a final concentration of 1 x 10(5) sperm/ml for 3 h, all oocytes were cultured in vitro for 52 h. Day-2 (52 h after insemination) embryos were classified according to their cleaved stages into 2-cell, 3- to 4-cell, 5- to 8-cell, and >8-cell stages; these were cultured separately for additional 4 days (Day 6). The resultant Day-6 blastocysts were classified according to the morphological diameter into 3 grades: Grade A, expanded blastocysts; Grade B, expanding blastocysts; and Grade C, early blastocysts. They were then analyzed chromosomally. The 3- to 4-cell and 5- to 8-cell embryos had significantly high blastocyst development rates (46.1 and 36.9%, respectively), and these blastocysts contained significantly more cells (40.2 and 42.4 cells, respectively) than those derived from 2-cell embryos and >8-cell embryos (28.6 and 26.5 cells, respectively). The incidence of chromosomal abnormalities was significantly higher in the blastocysts derived from 2-cell and >8-cell stage embryos than in the blastocysts derived from the other stage embryos. Furthermore, the grade A blastocysts had the lowest incidence of chromosomal abnormalities (35.3%) and contained the most cells (48.7 cells). Porcine in vitro production (IVP) yielded a high blastocyst rate and an excellent embryo quality when 3- to 4-cell and 5- to 8-cell stage embryos were selected on Day 2 after insemination. The same criteria yielded a higher quality of expanded blastocysts based on the stage of embryo development and morphology.     

核移植前激活受体卵子对牛核移植卵体外发育的影响

本研究探讨了核移植前对受体卵子进行激活、细胞融合开始时间及供体受精卵细胞周期调节对核移植卵体外发育的影响。其结果显示核移植前对受体卵子激活组的细胞融合率与对照组没有差异 ,但重组胚胎的卵裂率、8～ 16细胞期胚胎及囊胚的发育率比对照组明显提高 ;核移植前激活的受体卵子分别在卵子体外成熟开始的第30h和 4 5h与供体细胞进行细胞融合 ,结果 ,30h组的细胞融合率和卵裂率与 4 5h组没有差异 ,但发育到 8～ 16细胞期及囊胚的发育率均比 4 5h的高 ;将供体受精卵用诺考达唑 (Nocodazole)处理后 ,进行核移植的结果 ,处理组的细胞融合率、卵裂率、发育到 8～ 16细胞期和囊胚的发育率与对照组无差异     

Parthenogenetic activation and subsequent development of porcine oocytes activated by a combined electric pulse and butyrolactone I treatment

This study was designed to evaluate the parthenogenetic activation of porcine oocytes matured in vitro for a varied period after combined electric pulse (EP; 1500 V/cm, 100 microsec) and Butyrolactone I (BL I). After 36 h of maturation culture, the rates of activated oocytes and oocytes with two pronuclei were significantly lower than those of oocytes cultured for 42 and 48 h after EP. However, when treated by a combined EP and BL I (150 microM), these rates increased to the same level as 42 and 48 h oocytes. When oocytes cultured for 48 h and activated by a combined EP and BL I treatment were subsequently cultured in mNCSU37 medium, the rates of embryos cleaved and developed to the blastocyst stage were significantly higher than those in Whitten's medium. In contrast, when activated oocytes were cultured in mNCSU37 medium under two oxygen environments (5% vs 20% O(2)), there was no difference in the rates of cleavage, blastocyst formation and nuclear numbers per blastocyst. Our results demonstrated that the combined EP and BL I treatment of porcine oocytes matured in vitro is capable of producing high rates of good quality blastocysts when cultured in a suitable in vitro condition.     

小鼠4、8-细胞胚胎单卵裂球体外培养体系的研究

以CZB为基础培养液,培养小鼠4、8-细胞胚胎单卵裂球,研究葡萄糖、牛磺酸和猪输卵管上皮细胞共培养在其体外发育中的作用。结果表明:牛磺酸添加与否对4、8-细胞胚胎单卵裂球的囊胚发育率分别为29%、30%和14%、14%,无显著性差异(P>0.05)。添加葡萄糖后,4、8-细胞胚胎单卵裂球的囊胚发育率分别为36%和19%,有显著提高(P<0.05)。含有葡萄糖而牛磺酸的添加与否对4、8-细胞胚胎单卵裂球的囊胚发育率分别为36%、38%和19%、20%,无显著性差异(P>0.05)。各组内4、8-细胞胚胎单卵裂球形成的囊胚细胞数分别为(10.44±1.24～(12.43±1.18)和(7.57±0.97)～(8.48±1.16),均无显著性差异(P>0.05)。4、8-细胞胚胎单卵裂球与猪输卵管上皮细胞共培养,其囊胚发育率分别为47%和26%,囊胚细胞数分别为(17.57±1.13)和(11.43±0.92),均高于单一培养(P<0.05)。