Cumulus Oophorus Expansion of Bovine Oocytes Cultured in vitro: A SEM and TEM Study     

J. Laurincík  J. Pivko  P. Kroslák 《Reproduction in domestic animals》1992,27(4):217-228

Bovine cumulus-oocyte complexes (COCs) were aspirated from antral follicles (3–6 mm in diameter). COCs with a compact multilayered cumulus investment were cultured for 9, 13 and 24 hours, respectively, and processed for scanning or transmission electron microscopy after different periods of culture including a 0 hour control group. In all 0 hour control COCs initial formation of cumulus cell projections, blebs and microvilli were observed. The start of cumulus-oocyte disconnection in cattle was observed after 9 hours of in vitro culture. These connections were almost completely discrupted after 13 hours, when a continuous production of extracellular matrix was observed. The full expansion of corona radiata cells was not observed even after 24 hours. Some cumulus oophorus cells were bound together with junctions of the zonula adherens type after 24 hours when extracellular matrix was found only in deeper layers. The full expansion of corona radiata cells was not the prerequisite for disconnection of cumulus-oocyte complex. Inhalt: Kumulus oophorus Expansion von in vitro kultivierten Rinderoozyten: Eine SEM und TEM Untersuchung Kumulus-Oozyten-Komplexe (COC) von Rindern wurden aus antralen Follikeln aspiriert (Durchmesser 3–6 mm). COC's mit kompakten mehrschichtigen Kumulus wurden für 9, 13 und 24 Stunden kultiviert und für die Scanner- oder Transmissionselektronenmikroskopie vorbereitet. Verschiedene Kulturzeiten incl. einer 0-Stundengruppe als Kontrolle wurden untersucht. In allen Kontroll COC's wurden beginnende Formationen yon Kumuluszellprojektionen, Bläschen und Mikrovilli beobachtet. Nach 9-stündiger in vitro Kultur begann die Lösung zwischen Kumulus und Oozyte. Die Verbindungen waren nach 13 Stunden vollständig unterbrochen, wobei eine kontinuierliche Produktion extrazellulärer Matrix beobachtet wurde. Eine vollständige Expansion der Corona radiata Zellen wurde auch nicht nach 24 Stunden beobachtet. Nach 24 Stunden waren einige Cumulus Oophorus Zellen über Verbindungen des Zonula adherens Typs verknüpft, wobei extrazelluläre Matrix nur in den tieferen Zellschichten gefunden wurde. Eine vollständige Kumulusexpansion ist keine Voraussetzung für die Auflösung des Kumulus-Oozyten-Komplexes.  相似文献   

An improved vitrification protocol for equine immature oocytes,resulting in a first live foal          下载免费PDF全文

N. Ortiz‐Escribano  O. Bogado Pascottini  H. Woelders  L. Vandenberghe  C. De Schauwer  J. Govaere  E. Van den Abbeel  T. Vullers  C. Ververs  K. Roels  M. Van De Velde  A. Van Soom  K. Smits 《Equine veterinary journal》2018,50(3):391-397

  相似文献   

Differential effects of follistatin on porcine oocyte competence and cumulus cell gene expression in vitro          下载免费PDF全文

Z Guo  D Liu  G Liu  L Lv  Y Yang  B Fu  L Wang  Z Liu  H He  H Wu 《Reproduction in domestic animals》2018,53(1):3-10

  相似文献   

Effect Comparison of Adenovirus Vector Transfer into Different Buffalo Cells     

DUAN An-qin  PANG Chun-ying  ZHU Peng  LU Xing-rong  DENG Ting-xian  LIANG Xian-wei 《中国畜牧兽医》2016,43(10):2661-2665

Buffalo mammary epithelial cell,cumulus cell and fibroblasts were transfected by adenovirus vectors and compared their transfection efficiency.293 cells were transfected with pBHGloxdelE13cre and pDC316-eGFP by liposome,the virus was collected and titer was detected.Buffalo mammary epithelial cell,cumulus cell and fibroblasts were exposed to different multiplicity of infection (MOI) of adenovirus vectors.After 72 h,the cells were observed with inverted fluorescence microscope,and transfection efficiency was calculated.When the MOI was 25,50,100,200 and 400,the transfection efficiency of fibroblasts were 0.7%,7.0%,9.0%,12.5% and 34.0%,the transfection efficiency of cumulus cells were 42.5%,55.3%,57.4%,76.0% and 80.0%,the transfection efficiency of mammary epithelial cells were 88.7%,100%,100%,100% and 100%.The results showed that the transfection efficiency of mammary epithelial cell was the best,followed by cumulus cell,and fibroblast was poor.  相似文献   

单宁酸对猪卵母细胞体外成熟及胚胎发育能力的影响     

孙婧陶  姜超前  刘佳慧  张弛  王传玥  边雅  姜希卿  张琪  金君学 《中国畜牧兽医》2021,48(10):3708-3714

研究旨在探讨单宁酸对猪卵母细胞体外成熟质量及其胚胎发育能力的影响。在猪卵丘卵母细胞复合体（COCs）体外成熟培养液中添加不同浓度（0、1、10、100 μg/mL）单宁酸培养42 h后,检测COCs的扩散程度和卵丘细胞扩散指数,统计COCs的体外成熟率,检测成熟卵母细胞内谷胱甘肽（glutathione,GSH）、活性氧（reactive oxygen species,ROS）和生长分化因子9（growth differentiation factor 9,GDF9）的水平,并统计孤雌激活及体外受精胚胎48和168 h的卵裂率、囊胚率及囊胚总细胞数。结果显示,与对照组相比,10 μg/mL单宁酸组卵丘细胞扩散指数显著提高（P<0.05）,100 μg/mL单宁酸组显著降低（P<0.05）;1和10 μg/mL单宁酸组卵母细胞成熟率差异不显著（P>0.05）,100 μg/mL单宁酸组卵母细胞成熟率显著降低（P<0.05）;1和10 μg/mL单宁酸组GSH和GDF9水平显著提高（P<0.05）,ROS水平显著降低（P<0.05）。孤雌胚胎和体外受精胚胎发育能力结果显示,与对照组相比,各单宁酸组卵裂率差异不显著（P>0.05）,10 μg/mL单宁酸组孤雌胚胎囊胚率及体外受精胚胎囊胚率显著提高（P<0.05）,100 μg/mL单宁酸组孤雌胚胎囊胚细胞数及体外受精胚胎囊胚细胞数均显著低于其他各组（P<0.05）。以上结果表明,10 μg/mL单宁酸可通过提高卵丘细胞扩散能力及GSH和GDF9水平、降低卵母细胞内ROS水平,改善猪卵母细胞成熟质量,提高孤雌胚胎及体外受精胚胎的发育能力。  相似文献   

颗粒细胞单层对黄牛孤雌胚胎体外发育的影响     

彭辉  卿素珠  李莉  肖红卫  乔宪凤  郑新民 《湖北农业科学》2009,48(4)

为优化颗粒细胞单层共培养体系,研究了不同类型、不同种属的颗粒细胞单层及不同时间更换培养单层对黄牛孤雌胚胎体外发育的影响.结果表明.就卵裂率、囊胚率和囊胚孵出率而言.壁颗粒细胞单层组与丘颗粒细胞单层组之间无显著差异(P>0.05);来自黄牛、猪和小鼠的颗粒细胞单层在支持黄牛孤雌胚胎体外发育方面无显著差异(P>0.05):就囊胚率而言,共培养的第3天和第6天两次更换单层分别与共培养的第4天更换单层和始终不更换单层的对照组之间无显著差异(P>0.05),但第4天更换单层与对照组之间有显著差异(P<0.05).所以.不同类型和不同种属的颗粒细胞单层都能很好地支持黄牛孤雌胚胎的体外发育.在胚胎的体外发育过程中更换培养单层可以明显提高胚胎的囊胚率和囊胚孵出率,并且在共培养的第4天更换培养单层可以得到较高的囊胚率.  相似文献   

Role of oocyte‐derived paracrine factors in follicular development     

Chihiro Emori  Koji Sugiura 《Animal Science Journal》2014,85(6):627-633

Mammalian oocytes secrete transforming growth factor β (TGF‐β) superfamily proteins, such as growth differentiation factor 9 (GDF9), bone morphogenetic protein 6 (BMP6) and BMP15, and fibroblast growth factors (FGFs). These oocyte‐derived paracrine factors (ODPFs) play essential roles in regulating the differentiation and function of somatic granulosa cells as well as the development of ovarian follicles. In addition to the importance of individual ODPFs, emerging evidence suggests that the interaction of ODPF signals with other intra‐follicular signals, such as estrogen, is critical for folliculogenesis. In this review, we will discuss the current understanding of the role of ODPFs in follicular development with an emphasis on their interaction with estrogen signaling in regulation of the differentiation and function of granulosa cells.  相似文献   

卵丘细胞对牛卵母细胞体外成熟和体外受精的影响     

杨膺白  卢克焕 《广西农业生物科学》1994,(1)

本试验探讨了卵母细胞周围不同类型的卵丘细胞对牛卵母细胞体外成熟、受精及其随后的胚胎发育的影响。卵母细胞周围一般都含有多层卵丘细胞。按卵丘细胞的层数及形态将卵母细胞大致分为５类：（１）无卵丘细胞；（２）有２～３层卵丘细胞；（３）有４～５层卵丘细胞；（４）有６层以上卵丘细胞；（５）异常者（即卵母细胞外周有较厚的透明胶状物而无正常卵丘细胞）。以上５种卵母细胞经体外成熟培养并受精后，其卵裂率（分别为：４８．８％，７０．９％，８４．４％，８２．１％，６８．２％）及囊胚发育率（分别为：０．０％，１７．８％，３３．３％，５４．６％，２５．０％）除异常者外均随着卵丘细胞层数的增加而提高（Ｐ＜０．０５）。对含有２～６层卵丘细胞的卵母细胞成熟后进行不同程度的剥离处理。然后按剥离程度分为３组：（１）全部剥离；（２）仅剩放射冠；（３）放射冠外有２～３层卵丘细胞。受精后３组间的卵裂率无显著差异（分别为：８１．７％，８５．２％，８４．４％）。而囊胚发育率（分别为：１６．８％，２３．８％，２３．４％），全部剥离组明显低于其他两组（Ｐ＜０．０５）。试验结果表明卵丘细胞对卵母细胞的体外成熟有促进作用，从而影响随后的受精及胚胎发育。  相似文献   

Mouse embryo co‐culture with autologous cumulus cells and fetal development post‐embryo transfer     

Farha Naomi OMAR FAROUK  David STOTT  Marcela VLAD 《Animal Science Journal》2011,82(3):420-427

This study was conducted to examine the potential for implantation and sustainable fetal development of mouse embryos cultured from the pronuclear to blastocyst stage. Pronuclear embryos from ICR mice (Harlan Sprague‐Dawley) were cultured in Sydney IVF sequential media (Cook) to the blastocyst stage in medium only or co‐cultured with autologous cumulus cells. We also experimented with co‐culture in 100 µL drops. Drop co‐culture produced blastocyst formation rates with a mean of 47.0%, which was significantly higher (P < 0.05) compared to embryos cultured in identical culture conditions except without cumulus cells at 27.3%. Blastocysts obtained in vitro in Cook medium only and co‐cultured in Cook medium with cumulus cells were transferred to pseudopregnant females of ICR strain. The day of blastocyst transfer into surrogate females was designated as post‐transfer of blastocyst day 1 (PT 1). The implantation and fetal development was compared to embryo transfer of in vivo derived blastocysts, which served as controls. There were no statistical differences for implantation and fetal development rates for blastocysts cultured in vitro in either Cook medium only or co‐culture in Cook medium with cumulus cells compared to in vivo‐derived blastocysts. The advantage of the co‐culture system is in generating more blastocysts available for transfer.  相似文献   

Oocyte Transfer as an Analytical Tool:Vintage Experiments in Domestic Farm Animals     

R. H. F. Hunter  Xihe Li 《畜牧与生物技术杂志(英文版)》2011,2(4):224-229

Four transplant studies are described that focus on fertilisation and early development or the progression of unfertilised oocytes (eggs) in the oviduct.(1) Pig eggs transplanted from ovulations induce...  相似文献