Vitrification of mouse two-cell and blastocyst stage embryos in simplified closed system using either a hemi-straw or a hollow fiber device

Two-cell stage and blastocyst stage mouse embryos were equilibrated in a medium containing 7.5% ethylene glycol (EG) and 7.5% dimethyl sulfoxide (DMSO) for 8–15 min. Vitrification was performed in a medium containing 0.5 M sucrose and either 15% EG + 15% DMSO, 17.5% EG + 17.5% DMSO, or 20% EG + 20% DMSO for 30 s. They were then placed either on a hemi-straw (HS) or a hollow fiber vitrification (HFV) device and vitrified by cooled air inside a 0.5-ml straw. In two-cell embryos, a 100% survival rate was obtained from all groups except the 20% HS group (P > .05). All vitrified two-cell groups showed similar rates of blastocyst development to that of fresh control group (P > .05), except 17.5% and 20% HFV groups, which were significantly lower than the other groups (P < .05). In the blastocyst embryos, the HFV groups were divided into two subgroups (non-collapsed; HFV-NC and collapsed; HFV-C blastocyst). Re-expansion rate in 15% HFV-NC, 17.5% HFV-NC, and 15% HFV-C groups was reduced (P < .05), whereas the rest were similar to control. In conclusion, we established a simplified, reliable, and closed system for HFV vitrification applying hemi-straw, which does not require skilled practitioners.     

不同浓度CO2对牛卵母细胞体外成熟和受精的影响

对牛卵巢卵泡内卵母细胞在不同浓度的CO2培养箱内体外成熟、体外受精后的卵裂率及胚胎体外发育进行了研究。结果显示,含5%小牛血清(CS)的TCM-199内的卵母细胞在2%CO2条件下培养20h后,达到第二次减数分裂期(MⅡ期)的卵子数明显高于在5%CO2条件下培养的卵母细胞数(P<0.05)。两种浓度的CO2条件下培养的牛卵母细胞体外受精后的卵裂率无明显差异,2%CO2浓度下培养的卵母细胞的囊胚发育率高于5%CO2浓度下培养的卵母细胞。     

猪原始生殖嵴细胞(PGCs)建系因素的研究

从五指山猪(WSZP)近交系第8～13代培育群中，先后选用21头5～10月龄青年母猪，分别于授精后25～30d采集胎儿106个，进行原始生殖嵴(PGCs)细胞分离、培养等建系技术研究。以DMEM F10(1:1)为基础培养液，按添加或不添加生长因子，将培养液分为A、B、C3种，并以STO细胞作饲养层，在38℃、5.0％CO2和湿润的气相中进行培养建系。结果获得胚胎生殖嵴细胞(EG)细胞系6个细胞株，其中1个EG细胞株传至11代、2个传至5代、1个传至4代、2个传至3代冻存。并进行了AKP染色、体外分化、冷冻-解冻复苏和嵌合体制作等鉴定研究。研究发现：不同胚龄对EG细胞建系具有一定影响，不同培养液对EG细胞建系效果不同，STO细胞饲养层的质量是建株、传代、冷冻-解冻复苏的关键因素之一。EG细胞系的初步建立，为今后筛选进入种系的EG细胞系、实施体外基因操作提供了可能。     

大鼠心肌条件培养基对ICR小鼠胚胎干细胞分离克隆的影响

采用大鼠心肌条件培养基(RH CM)培养ICR小鼠的桑椹胚和囊胚,发现由囊胚分离的ES细胞传代后ES集落的出现率显著高于桑椹胚(P<0.05),囊胚更适合作为ES细胞分离克隆的材料。以RH CM为培养基的试验组ES细胞传代的平均时间间隔为38 h,对照组传代的时间间隔平均为78 h,两者差异显著(P<0.05)。表明RH CM能够促进ES细胞贴壁增殖和ES集落的形成,有效地维持ES细胞未分化状态。试验中设计的3 种培养条件对原代ES集落的形成影响不显著,但对传代后的ES集落的形成和传代的代次有显著差异。其中以MEF作饲养层,添加RH CM培养基的效果最好。     

烟青虫初孵幼虫的高效快速收集方法

比较了常用的烟青虫人工收集方法和新研发的烟青虫初孵幼虫快速收集方法的效果。试验结果表明,与人工收集方法相比,快速收集方法获得初孵幼虫所用时间少,收集的初孵幼虫存活率搞,达96.33%,且收集的幼虫发育更加整齐,雌、雄蛹单头质量(雌蛹295.33 mg,雄蛹284.66 mg)略大;还可节省大量人力,避免了人工挑虫对初孵幼虫的伤害,保障了烟青虫实验种群的大量优质扩繁。     

A comparison of the fatty acid profiles of adult tissues,and newly hatched,fed and starved Lysmata amboinensis larvae

This study evaluated the lipid content and fatty acid (FA) profile of the hepatopancreas, ovaries and tail muscle of Lysmata amboinensis broodstock, as well as newly hatched larvae subjected to a period of starvation or feed from hatch to Zoea 2. The hepatopancreas had a high lipid content, confirming its role as a process and storage organ in L. amboinensis. Lipids were also a major component of ovarian dry weight, in agreement with reports on other crustaceans during maturation. The tail muscle, being a functional rather than a storage organ, contained low total lipids and was the tissue that closely resembled the FA profile of the newly hatched larvae. Saturated fatty acids (SFAs) and highly unsaturated fatty acids (HUFAs) were the most abundant components of the lipid profiles in broodstock and larvae. The HUFAs docosahexaenoic and eicosapentaenoic were preferentially retained during nutritional stress, confirming their importance for marine cleaner shrimp during early larval development. It appeared polyunsaturated fatty acid and HUFA requirements were met through the larval diet. The SFAs stearic and palmitic were abundant in adult tissues and larvae, whereas monounsaturated fatty acids may have been preferentially catabolized to meet energetic and metabolic larval requirements.     

牛IVF性控切割胚胎和整胚的冷冻保存研究

本研究分3个实验对比了采用不同处理的常规冷冻和玻璃化冷冻保存358枚体外受精(IVF)性控切割囊胚和326枚整胚的保存效果。结果表明：(1)用于冷冻整胚的常规方法对冷冻性控切割胚胎同样有效；(2)利用含10％EG 10％FBS的DPBS冷冻牛IVF控切割胚胎或整胚的效果明显优于用含10％甘油 0.4%BSA或1．4mol／LEG 0．1％PVOH的DMPBS；(3)采用微玻管玻璃化冷冻保存牛IVF性控切割胚胎的效果显著优于常规冷冻法。     

澳洲淡水龙虾受精卵到仔虾营养物质变化分析

摘要：为探究澳洲淡水龙虾受精卵发育到仔虾过程中营养物质变化情况,本研究分别对澳洲淡水龙虾前无节幼体期、复眼色素形成期、孵化准备期受精卵及初孵仔虾的营养成分组成进行了系统性分析,研究表明：水分含量随着受精卵逐渐发育成仔虾而显著升高(P<0.05),粗蛋白和粗脂肪含量则随发育过程而显著降低(P<0.05),灰分含量以初孵仔虾最高(P<0.05);总氨基酸(TAA)、必需氨基酸(EAA)和呈味氨基酸(DAA)均随发育过程而显著降低(P<0.05);在AAS模式下,受精卵和初孵仔虾第一限制性氨基酸均为Met+Cys,受精卵第二限制性氨基酸为Leu,初孵仔虾第二限制性氨基酸为Ile。在CS模式下,受精卵和初孵仔虾第一限制性氨基酸也是Met+Cys,前无节幼体期受精卵第二限制性氨基酸为Val,复眼色素形成期之后的受精卵和初孵仔虾第二限制性氨基酸均为Ile。受精卵和初孵仔虾的EAAI随着受精卵发育到初孵仔虾而显著降低(P<0.05)。澳洲淡水龙虾受精卵和初孵仔虾共测出主要脂肪酸10种,初孵仔虾的PUFA及DHA含量显著低于受精卵(P<0.05),EPA+DHA含量却显著高于受精卵(P<0.05)。综上,澳洲淡水龙虾初孵仔虾灰分及EPA+DHA含量显著高于受精卵,故在澳洲淡水龙虾仔虾开口饵料配制中可考虑适当提高磷酸二氢钙及EPA+DHA添加量。     

关于桑蚕(Bombyx mori L.)对人工饮料摄食性的研究──Ⅰ.不同收蚁时间和饲料含水率对小蚕发育的影响

为解决发育不齐的问题，就家蚕不同收蚁时间、不同饲料含水率对家蚕幼虫发育的影响进行了研究。结果发现：①发育整齐度，孵化当天收蚁＞二夜包＞三夜包＞四夜包；②饲料含水率以73％左右为适当．过高或过低都会延缓蚕的发育，影响发育整齐度。     

催青环境及分段收蚁对原蚕性别调控研究

试验结果表明 ,通过改变催青环境和分段收蚁可以人工调控家蚕的性别比例。在原种催青期中高温干燥处理 ,可使原蚕品种 872和鲁玉 蚕比例分别提高 1 3 %和 2 4% ,达极显著和显著水平 ;对原蚕 795和871品种分段收蚁 ,第 1天可提高♀性比例 1 0 %～ 2 0 % ,最后一天 (第 3天 )提高 蚕比例 ,795品种最高为2 8%～ 3 6% ,达极显著水平。