表皮生长因子对水牛胚胎体外发育及凋亡的影响

为了探讨表皮生长因子(EGF)对水牛早期胚胎体外发育及凋亡的影响,通过收集屠宰场卵巢卵母细胞进行体外成熟和体外受精,将假定的受精卵置于含不同浓度EGF(0,25,50和100 ng/mL)的培养液中培养,检查分裂率和囊胚发育率,用细胞凋亡试剂盒(Annexin-V-FluosStaining kit)试剂染色,统计囊胚细胞凋亡率和坏死率。结果表明:50 ng/mL EGF组的孵化囊胚率显著高于对照组(P<0.05),该组细胞凋亡率和坏死率显著低于对照组(P<0.05)。100 ng/mL EGF的卵裂率、囊胚率、D7囊胚率和孵化囊胚率显著低于对照组和其他试验组(P<0.05)。细胞凋亡率和坏死率显著高于其他各组(P<0.05)。提示:一定浓度的EGF可提高囊胚孵化率,并可抑制胚胎细胞的凋亡。     

小鼠卵母细胞透明带的切割

透明带的切割是哺乳动物早期胚胎显微操作技术中的一个重要环节，本文以小鼠为例介绍一种新的透明带切割方法。     

无透明带体细胞核移植研究进展

目前,体细胞核移植主要还是沿用Willadsen发明的显微操作的去核、注核、融合和激活来完成。由于其核移植效率较低,且对仪器设备和技术人员的要求较高,因此,简化并建立一套更适合于生产条件下的体细胞核移植程序,从而提高核移植效率一直是人们所不懈努力的。作者就无透明带核移植技术和单个胚胎培养系统作一介绍。     

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。     

3种中药有效成分对小鼠胚胎体外发育的影响

以mCZB添加抗生素为对照组,以分别添加0.1 mg/L小檗碱(berberine,BR)、0.5 mg/L川芎嗪(ligustrazine,Lig)和4 mg/L黄芩苷(baicalin,Bai)为试验组,比较各组胚胎孵化率、微滴中一氧化氮(Nitric oxide,NO)和丙二醛(malondialdehyde,MDA)含量及孵化胚胎的细胞数量,从而探讨中药有效成分对小鼠早期胚胎体外发育的影响。结果显示,Bai组(80.2%,81.3%)、Lig组(78.9%,77.6%)及BR组(76.7%,71.6%)胚胎孵化率极显著高于对照组(50.7%、47.8%)(P<0.01)。各组间NO含量,72 h较24 h均有减少,而120 h又均有增加且最高。各组间MDA含量,72 h和120 h与24 h比较呈现增加趋势,除Bai组120 h外,对照组均高于试验组。孵化胚胎细胞计数结果显示,BR组(87.2±8.6)、Lig组(83.9±7.7)和Bai组(81.9±6.2)与对照组(77.4±5.6)差异极显著(P<0.01)。结果表明:3种中药有效成分均能促进小鼠早期胚胎的体外发育及胚胎细胞增殖,并对NO和MDA含量有一定影响。     

氯化锶对去透明带卵母细胞孤雌激活效果及孤雌胚胎体外发育体系的研究

本实验以昆明小鼠为研究对象,探讨了5 mmoL/L、10 mmoL/L及20 mmoL/L氯化锶(SrCl2)激活去透明带MⅡ期卵母细胞的效果,并比较了孤雌激活卵母细胞在普通DMEM培养液与添加有10 ng/mL有白血病抑制因子(LIF)的DMEM培养液中的体外发育率,旨在探讨去透明带卵母细胞适宜的SrCl2孤雌激活浓度及孤雌胚胎体外培养体系。结果表明:10 mmoL/L与5 mmoL/L SrCl2作用6 h所得激活率差异不显著(46.00%vs 40.38%,P>0.05),但显著高于20 mmoL/L SrCl2的激活率(46.00%vs 24.62%,P<0.05);将10 mmoL/L SrCl2激活处理6 h后得到的激活胚胎分别以DMEM培养液和LIF+DMEM培养液培养48 h,其桑葚胚分化率分别为44.00%及84.62%,组间差异显著(P<0.05)。因此,宜采用10 mmoL/L SrCl2激活去透明带MⅡ期卵母细胞,并移入到10 ng/mL LIF+DMEM培养液进行体外培养。     

表皮生长因子对母畜生殖的影响

表皮生长因子（ＥＧＦ）是一种具有广泛生物学效应的单链多肽。家畜多种组织中都含有ＥＧＦ及其受体，在生殖系统中亦同样存在。因此ＥＧＦ在家畜生殖中所起的作用愈来愈被人们所重视。它可通过自分泌、旁分泌及内分泌途径对生殖器官及生殖激素产生调控作用，在性腺功能、卵泡生长、子宫功能及胚胎发育等方面都起重要的作用。     

玻璃化冷冻对小鼠卵母细胞透明带超微结构变化及体外受精效果的影响

本实验采用OPS法玻璃化冷冻小鼠卵母细胞,通过扫描电镜观察透明带表面物理特性的变化。旨在研究冷冻导致小鼠卵母细胞透明带表面超微结构的变化以及冷冻卵母细胞体外受精后胚胎发育率和精子穿透透明带能力的影响。结果表明:冷冻后透明带表面网状结构消退或消失,发生熔融样变化。冷冻组中A型透明带比例(57.14%)显著低于毒性实验组(79.32%)及对照组(81.25%)(P<0.05),而发生熔融样变化的比例显著高于其他两组(P<0.05)。体外受精后,冷冻组的卵裂率和囊胚发育率(60.90%,39.51%)均显著低于毒性实验组及对照组(81.74%,55.72%;87.47%,61.63%)。体外受精4 h,冷冻组透明带平均结合精子数(10.56)与毒性实验组和对照组(10.26,10.21)间均无显著性差异(P>0.05);但受精后对透明带消化处理发现,冷冻组紧密结合精子的平均个数(3.28)与其他两组(5.11,5.21)存在显著性差异(P<0.05)。结果显示,冷冻导致透明带表面物理特性的变化,进而造成受精率下降。     

猪卵母细胞和早期胚胎透明带杨氏模量变化

在39℃环境下,利用微管吸吮的方法,在倒置显微镜下,通过显微操作装置用显微玻璃管吸吮猪卵母细胞和早期胚胎来测定其透明带的杨氏模量。结果显示,从猪卵巢抽取的未经体外成熟培养的卵母细胞(GV期)一直到早期囊胚(未扩张囊胚)阶段,其透明带杨氏模量一直都无显著变化,均集中在20 kPa左右,但当发育到扩张囊胚期其透明带杨氏模量明显变大(P0.05),且扩张囊胚中化学激活的明显大于体外受精的(P0.05)。结果提示,在猪卵母细胞和早期胚胎发育进程中,其透明带的刚性在扩张囊胚期最大,这为通过某些理化方法来帮助扩张囊胚期的胚胎孵化,从而提高囊胚的孵化率并最终提高妊娠率提供了力学方面的理论参考。     

Effect of zearalenone on the growth of mouse embryos from blastocysts to the egg cylinder stage in vitro

Embryos were harvested at the blastocyst stage from nontreated outbred mice and were grown in vitro for 4 days. Embryos cultured in control medium hatched and grew to the egg cylinder stage. Purified zearalenone (ZEN) added to the culture medium at concentrations of 8.5 to 68 micrograms/ml decreased the number of embryos growing, with a 50% decrease in the number growing in 32 micrograms of ZEN/ml of medium. Embryos growing in ZEN had decreased numbers of cells derived from the inner cell mass, normal growth of the trophoblast, less cellular differentiation than was seen in control embryos, and increased numbers of phagosomes. Undifferentiated cells of the inner cell mass of control and treated embryos were of the same size, as determined by morphometric analysis. Addition of 25 micrograms of estradiol/ml of culture medium caused no decrease in number of embryos growing or in embryonic size. Saturation of culture medium with ZEN (68 micrograms/ml) did not inhibit the growth of a tissue culture line of goat synovial cells. Seemingly, ZEN at concentrations near saturation inhibited the growth of mouse embryos in vitro. This effect was not duplicated with similar concentrations of estradiol and was not manifested in culture-adapted cells.     

Effect of epidermal growth factor on healing of corneal endothelial cells in cats

Epidermal growth factor was injected intracamerally into the anterior chamber of the right eye of 9 cats. The central portion of the cornea in 8 of the 9 cats that had been cryoinjured. Effect of epidermal growth factor on the repair of endothelial cells in cats was evaluated by endothelial specular microscopy. Endothelial cell density and corneal thickness were studied quantitatively, as a measure of endothelial cell function. The repair process also was evaluated qualitatively by studying morphologic changes, developing as a result of reendothelialization and return to normal function. Seemingly, differences between rate of healing of cryoinjured eyes injected with epidermal growth factor and that in nontreated eyes were not significant (P = 0.86). The endothelial repair process was characterized by enlargement and migration of adjacent noninjured cells.     

生长因子在着床前鼠胚发育中的作用

近年来,大量的研究显示。生长因子在哺乳动物着床前胚胎的发育中起了十分重要的作用。首先。哺乳动物着床前胚胎以及生殖道中可以表达多种生长因子及其受体,并且在体外实验中通过添加生长因子,如EGF、IGF-Ⅰ、IGF-Ⅱ等能显著促进胚胎细胞的增生与分化。同时,如果加入抗生长因子抗体或受体抑制剂。可以阻     

Effect of insulin,transferrin and selenium and epidermal growth factor on development of buffalo oocytes to the blastocyst stage in vitro in serum-free,semidefined media

The in vitro development of buffalo oocytes up to the blastocyst stage was studied in serum-free, semidefined media containing bovine serum albumin, follicle-stimulating hormone (FSH), insulin, transferrin and selenium (ITS) and epidermal growth factor (EGF). In experiment 1, oocytes aspirated from abattoir-derived ovaries were cultured in eight serum-free, semidefined culture media containing different combinations of these four factors. In experiment 2, the maturation of buffalo oocytes and the development of the embryos were compared in a complex co-culture system and in the serum-free, semidefined media. Supplementation with FSH and EGF significantly (P < 0.05) increased the maturation rates of buffalo oocytes, and the yield of blastocysts was higher (P < 0.05) in media containing EGF and ITS. The yield of blastocysts was lower in the serum-free semidefined media (P < 0.05) than in the complex co-culture system.     

Effects of epidermal growth factor (EGF) on fetal islet B cells in vitro

ABSTRACT. It has been reported that when the rat fetus is treated with streptozotocin (STZ) in vivo, islet B cells are destroyed but later recover. To investigate the process of the recovery of B cells after in vitro treatment of the fetal pancreas with STZ and the role of epidermal growth factor (EGF) in the recovery of B cells, we measured the level of insulin released from the cultured fetal pancreas and examined it histologically. As a result, we immunohistologically confirmed the regeneration of B cells in the pancreas that had been cultured for 48 hr after destruction of islet B cells by STZ treatment. An immunohistologic study using proliferating cell nuclear antigen (PCNA) showed that without the addition of EGF, the cell division index was significantly higher in the STZ-treated group (STZ group) than in the untreated group (intact group), whereas with the addition of EGF, the cell division index increased in both groups, but EGF did not have a significant cell division-promoting effect on the pancreas in the STZ group. The addition of EGF caused a significant decrease in the concentration of insulin in culture medium in both groups. These results indicate that EGF has a cell growth-promoting effect on intact fetal pancreas in vitro but has the effect of inhibiting the release of insulin, and thus suggest that EGF does not trigger the regeneration of islet B cells.     

Effect of phosphonoformic acid in the development of bovine embryos in vitro

This research evaluated the ability of phosphonoformic acid to inhibit bovine herpesvirus 1 (BHV-1) in cumulus cells commonly used in co-culture with bovine in vitro-produced embryos. At 200 and 400 microg/ml, phosphonoformic acid inhibited 4 logs of BHV-1. Subsequently, phosphonoformic acid (200 and 400 microg/ml) added to both in vitro fertilization and culture medium resulted in a decrease in the proportion of developed blastocysts, and the number of cells per blastocyst was lower in the treated embryos. Therefore, while phosphonoformic acid can effectively inhibit replication of BHV-1 in co-culture cells, it also inhibits development of in vitro-produced bovine embryos.     

氨基酸、维生素对水牛体外受精胚胎体外发育的影响

通过在培养液中添加不同浓度的氨基酸或维生素,探讨其对水牛体外受精(IVF)胚胎体外发育的影响.结果表明:(1)非必需氨基酸可显著提高水牛卵母细胞IVF后胚胎的分裂率,但对囊胚发育率无显著影响;(2)低浓度的必需氨基酸对水牛IVF胚胎的发育具有一定促进作用,但高浓度则有抑制作用;(3)维生素对水牛IVF胚胎发育则有促进作用.在培养液中添加维生素,水牛IVF胚胎的分裂率和第7天囊胚发育率显著提高.