鹌鹑胚胎的换壳培养

将25枚产后不到24 h的鹌鹑种蛋,作换壳培养试验。用25枚正常鹌鹑种蛋(对照组Ⅰ)和25枚鹌鹑无精蛋(对照组Ⅱ)作为对照,按常规方法孵化。结果:试验组有1羽鹌鹑正常出雏,孵化率迭4％。对照组Ⅰ有20羽鹌鹑孵出,孵化率为80％,对照组Ⅱ无鹌鹑孵出,无孤雌生殖现象发生。说明该试验具有可行性,它为今后实施胚胎移植和体外受精等试验作了准备。     

鸡胚胎体外操作技术的研究现状和进展

本文介绍了禽类胚胎操作技术方面的概况，对受精卵收集及体外受精、早期胚胎体外培养、人造材料蛋壳培养胚胎、代用蛋壳培养胚胎、嵌合体鸡的生产、转基因鸡生产等的进展及发展进行了综述。     

不同时期鸡胚胎体外培养的比较研究

试验采用代用蛋壳体外培养方法进行了不同时期鸡胚胎换蛋壳培养的研究。结果显示：体外受精卵体外培养发育率为10％。X期胚胎体外培养的孵化率为1．2％；孵化1日龄的胚胎体外培养的孵化率为7．9％；直接去1／4壳的鸡胚孵化率为3．7％；鸡胚胎换蛋壳以后，添加6～8札的稀蛋白对换壳鸡胚发育比较适宜。     

在代用蛋壳中孵化鸡胚

本实验将正常孵化７２ｈ的鸡胚转移到代用蛋壳内培养。鸡胚的存活率在第１０ｄ和第１８ｄ时分别是１００％和９０％（２３／２５）。孵化率为５２％（１３／２５）。     

动物胚胎体外培养研究进展

近年来动物胚胎体外培养取得了新进展.致密化前期胚胎利用葡萄糖作为能量底物的能力有限,但致密化后胚胎培养中葡萄糖的添加是必须的.自由氨基酸在早期胚胎的发育中是必需的,非必须氨基酸在致密化前发挥作用,致密化后期胚胎则需要所有氨基酸.血清对早期胚胎的发育有多重影响,BSA等一些生物大分子物质的配合添加可以取代血清的胚胎营养作用,并消除血清的负作用.在无血清培养液中,一些生长因子和细胞因子的加入是必须的.GSH等抗氧化物质添加进培养液可以对抗反应氧的作用,对胚胎发育起到保护作用.     

动物胚胎体外培养研究进展

近年来动物胚胎体培养取得了新进展，致密化前期胚胎利用葡萄糖作为能量底物的能力有限，但致密化后胚胎培养中葡萄糖的添加是必须的，由于氨基酸在早期胚胎的发育中是必需的，非必须氨基酸在致密化前发挥作用，致密化后期胚胎则需要所有氨基酸，血清对早期胚胎的发育有多重影响，BSA等一些生物大分子物质的配合添加可以取代血清的胚胎营养作用，并消除血清的负作用，在无血清培养液中，一些生长因子和细胞因子的加入是必须的，GSH等抗氧化物质添加进培养液可以对抗反应氧的作用，对胚胎发育起到保护作用。     

山羊早期胚胎体外培养试验

山羊早期胚胎体外培养试验邓世全（四川畜牧兽医学院荣昌６３２４６０）前言＊胚胎移植不但可提高优良母畜的繁殖潜力，扩大＊良种畜群，也是研究受精、胚胎发育和遗传工程的有力＊手段，还是体外受精、胚胎分割，核移植、嵌合体和基因＊导入技术的必要组成部分。我们在兔...     

牛体外受精胚胎的体外培养研究进展

家畜体外受精（IVF）技术自上世纪八十年代获得成功以来，发展十分迅速，九十年代已进入试管胚的开发应用阶段。目前，IVF胚胎的质量与体内胚相比仍有很大差距。在进行IVF胚胎体外培养时，常会出现“体外发育阻断”现象，主要表现为细胞数较少、卵裂球形状不规则、存在死亡的细胞和细胞质碎片、发育速率和抗冻性降低以及酶活性和营养物摄取的改变等方面。近年来，为了克服胚胎体外发育所存在的某些缺陷，国内、外对不同动物的胚胎体外培养体系和培养方法进行了系统研究，取得了长足进展。本文就牛IVF胚胎体外培养方面的研究进展综述如下。     

板蓝根多糖抗小鹅瘟病毒的作用

在鹅胚内用板蓝根多糖(IRPS)作直接杀毒和阻断感染试验,对鹅胚尿囊液中小鹅瘟病毒(GPV)含量、鹅胚存活时间、存活率、病变和免疫组化进行测定。结果显示,40μg/胚IRPS阻断感染组,GPV接种后72h ELISA检测为阳性的尿囊液最高稀释倍数23;鹅胚平均存活时间192h;存活率6/6;胚体肉眼观察未见病变;切片镜检可见肝、肾轻度淤血和细胞变性,心、脑轻度淤血和水肿;免疫组化显示肝、肾内GPV表达抗原为弱阳性,心、脑内为阴性。而攻毒对照组尿囊液GPV阳性的最高稀释倍数28;鹅胚存活时间138h;存活率0/6;胚体严重萎缩、出血、水肿,各组织细胞碎裂、坏死,组织内GPV表达抗原全为强阳性。两者比较,各指标具有极显著差异。这表明在鹅胚内IR-PS具有极强的阻断GPV感染、增殖和保护胚体组织器官免受其损伤的作用。     

Function of berberine on porcine in vitro fertilization embryo development and differential expression analysis of microRNAs

The effect of berberine (Ber) on in vitro fertilization (IVF) embryo development in pigs and the associated differential expression of microRNAs (miRNAs) in the embryo were investigated. NCSU‐23 embryonic culture medium was used for a control group, while NCSU‐23 embryonic culture medium added with Ber was used for a Ber group. The embryo development rates in these groups were determined, and the zygotes, 4‐ and 8‐cell embryos, and blastocysts were collected for cDNA microarray analysis. The development rates of 2‐, 4‐, 8‐cell embryos and blastocysts were significantly higher in the Ber group than those in the control group (p < 0.01). The differentially expressed miRNAs in the 8‐cell versus the 4‐cell stage in control group as well as in the 8‐cell Ber group versus the 8‐cell control group overlapped, and it was found that nine miRNAs were commonly upregulated and two of them were downregulated, while there was no overlap among the other groups. The target genes of Ber‐regulated miRNAs at the 8‐cell stage were mainly associated with the molecular pathway of nucleic acid and protein synthesis. These findings suggest that Ber may regulate the expression of miRNAs at the 8‐cell stage, which is beneficial to provide material reserves for the maternal to zygote transition of porcine embryos, thereby increasing the porcine IVF embryo development rate.     

鹅副粘病毒对鸡胚及雏鸡致病性的初步研究

鹅副粘病毒是一种引起多种家禽疾病的病毒,通过人工感染鸡胚和雏鸡试验,其结果表明本病毒主要以呼吸系统、消化系统病变为主要临床症状和特征,并且可导致10日龄的鸡胚100％死亡,对雏鸡的致死率为85％。因此,可见其致死率高,广大养殖户应重视鹅副粘病毒的预防。     

鹅盲肠微生物体外利用ADF/NDF的初步研究

以苜蓿草粉、黑麦草粉和混合草粉(质量比为1∶1)为底物,接种鹅盲肠食糜滤液进行体外培养。试验结果为:培养液pH值在0~3 h下降到6.9左右,后逐渐趋于稳定;苜蓿草粉ADF、NDF和半纤维素的消失率分别为1.15%,1.18%,-0.19%;黑麦草粉三者的消失率分别为1.45%,9.05%,3.07%;而混合草粉三者的消失率分别为15.72%,13.2%,9.87%。与单一草粉的ADF、NDF和半纤维素的消失率都有显著差异(P<0.05)。结果表明:在体外条件下,鹅盲肠微生物对不同来源的ADF、NDF和半纤维素有一定的利用能力,且对混合草粉ADF、NDF和半纤维素的利用能力强于单一草粉。     

鹅肝细胞的高纯分离及培养

采用简单的＂胶原酶直接消化法＂分离鹅胚胎肝细胞,得到的肝细胞纯度高,活性好;HE染色显示大量双核或多核细胞,细胞具有旺盛增殖分裂能力;肝脏特异性基因白蛋白基因检测显示所分离细胞为成熟肝脏细胞。该法操作简单、高效,是获得鹅原代肝细胞的一种较好的方法,有利于以鹅原代肝细胞为模型的体外研究工作的开展。     

The method of chicken whole embryo culture using the eggshell windowing,surrogate eggshell and ex ovo culture system

1. The unique accessibility of the avian embryo have made them an ideal model for the study of development and genome editing. Chicken whole embryo culture has provided important insights into toxicity tests, gene manipulation, clarifying gene functions, cell transplantation and cell tracking.

2. A simple technique for chicken manipulation is eggshell windowing, without or with seal, the latter having demonstrated some improvement in hatching rates.

3. Likewise, a surrogate eggshell system provides an accessible model for manipulation during chicken and quail development, with a higher hatchability compared to the simple windowing method.

4. The development of the chicken ex ovo culture systems in a synthetic environment as an efficient technique for imaging and microsurgery applications has enabled the study of important events of live chicken embryos at a specific time point.

5. This short review illustrates recent applications of well-designed whole embryo culture systems as a robust model for research into numerous biological mechanism, drug discovery, gene manipulating and production of functional proteins.     

第X期至“蚊虫珠”期鸡胚的易壳培养

将发育至第X期的鸡胚转和代用蛋壳内，分为4组分别添加蛋清至满或空余一定气室，正立或倒置孵化，观察鸡胚发育至“蚊虫珠”期胚胎的比较。结果发现鸡胚仅在代用蛋壳气室充盈无气泡、并与壳膜充分接触量，85%（34/40）的鸡胚能发育至“蚊虫珠”期。     

牛的体外受精技术研究进展

本文阐述了体外受精技术在牛胚胎工程和育种研究中的意义，介绍了牛体外受精技术的研究历史和目前的研究进展，对牛体外受精技术的技术指标现状及经济效益进行了评估，并就牛的体外受精技术当前研究存在的问题作了评述，对其前景做了展望。     

Effect of season and superstimulatory treatment on in vivo and in vitro embryo production in wood bison (Bison bison athabascae)

Two experiments were done using a two-by-two design to determine the effects of season and superstimulatory protocol on embryo production in wood bison. In Experiment 1 (in vivo-derived embryos), ovarian superstimulation was induced in female bison during the ovulatory and anovulatory seasons with either two or three doses of FSH given every-other-day (FSH × 2 vs. FSH × 3, respectively). Bison were given hCG to induce ovulation, inseminated 12 and 24 hr after hCG, and embryos were collected 8 days after hCG (n = 10 bison/group). In Experiment 2 (in vitro embryo production), ovarian superstimulation was induced in female bison during the ovulatory and anovulatory seasons with two doses of FSH, and in vivo maturation of the cumulus–oocyte complexes (COC) was induced with hCG at either 48 or 72 hr after the last dose of FSH. COC were collected 34 hr after hCG, and expanded COC were used for in vitro fertilization and culture. In Experiment 1, the number of follicles ≥9 mm, the proportion of follicles that ovulated, the number of CL, and the total number of ova/embryos collected did not differ between seasons or treatment groups, but the number of transferable embryos was greater (p < .05) in the ovulatory season. In Experiment 2, no differences were detected between seasons or treatment groups for any end point. The number of transferable embryos produced per bison was greatest (p < .05) using in vitro fertilization and was unaffected by season (1.5 ± 0.2 and 1.1 ± 0.3 during anovulatory and ovulatory seasons, respectively), in contrast to in vivo embryo production which was affected by season (0.1 ± 0.01 and 0.7 ± 0.2 during anovulatory and ovulatory seasons, respectively). Results demonstrate that transferable embryos can be produced throughout the year in wood bison by both in vivo and in vitro techniques, but the efficiency of embryo production of in vivo-derived embryos is significantly lower during the anovulatory season.