大鼠原代肝细胞的分离与培养

体外培养的大鼠原代肝细胞是研究药物代谢的有效手段,亦是细胞生物学和分子生物学的必需技术。常用的肝细胞分离方法有机械法和酶法。采用何种方法能够获取产率高、活性好、功能强的肝细胞,对于进一步的试验研究十分必要。随着科技的发展,肝细胞分离和培养技术在不断改进。在前     

成年鸡原代肝细胞两种分离方法的比较

为获得数量多及活力好的原代肝细胞,本试验分别采用原位二步法与半原位法分离鸡的肝细胞.应用MTT法比较了两种分离方法的细胞活力,应用电镜观察了分离细胞的超微结构,并分别在体外培养的第1、3、5天和第7天检测肝细胞上清液的AST、ALT、ALB和GGT,以鉴定鸡原代肝细胞的生化功能.结果显示,采用半原位法分离的鸡肝细胞与原位二步法相比,细胞数量多、活力高、结构完整,可在体外持续培养7d并稳定地保持肝细胞的形态与功能.     

鹅肝细胞的分离培养及其三酰甘油合成功能检测

本文旨在探讨鹅肝细胞的分离与原代培养方法,并对分离的肝细胞三酰甘油(TG)合成功能进行检测.采用改进的Seglen二步法分离纯化来自10日龄仔鹅肝脏的肝细胞,并进行原代培养.在倒置显微镜下观察肝细胞形态变化,MTT比色法测定细胞活性,用脂肪酸培养肝细胞并检测其形态、活性和细胞内TG含量的变化.结果显示,分离出的肝细胞形态完整、贴壁良好、活性高,细胞活性曲线与理论“S”型细胞生长曲线基本符合.另外,脂肪酸能增加肝细胞体积、活性和TG含量.结果表明,本研究成功分离了鹅肝细胞并进行了原代培养,肝细胞生酯功能正常.     

原代大鼠肝细胞分离培养方法改良

改进传统的两步原位胶原酶灌流法,调整灌流液及灌流条件,梯度离心纯化肝细胞.结果表明,分离出的大鼠肝细胞产量、纯度、活力均较高.该方法为一高效实用的原代大鼠肝细胞分离技术.     

大鼠肺微血管内皮细胞的体外分离培养与纯化

旨在建立一种简单易行、获得细胞纯度较高的体外分离、培养大鼠肺微血管内皮细胞的方法。取5~7日龄SD大鼠分离外周肺组织,组织植块法获得原代大鼠肺微血管内皮细胞,37℃、5%CO_2培养箱中传代培养,采用免疫磁珠法对其进行分离纯化,并对分离纯化后的细胞进行免疫荧光及流式细胞鉴定,通过MTT比色分析法准确、安全、可靠地检测纯化后传代肺微血管内皮细胞的增殖情况。结果显示:倒置显微镜下观察单个细胞呈短梭形或多角形,汇合后呈铺路石样,单层贴壁生长,细胞分离纯化后经免疫荧光检测CD31呈阳性,流式细胞仪检测淋巴管内皮细胞表面特有标记VEGFR-3呈阴性(P=0.1,且P0.5),且复苏后经MTT测得传代细胞的增殖情况良好。成功建立了一种分离高纯度大鼠肺微血管内皮细胞的体外培养方法。     

佐剂性关节炎大鼠成纤维样滑膜细胞的体外培养与鉴定

探讨佐剂性关节炎(AA)大鼠成纤维样滑膜细胞(FLS)的体外培养及鉴定方法,制备AA大鼠模型,采用组织块培养法培养AA大鼠FLS,并通过形态学、透射电镜及RT-PCR方法对FLS进行鉴定.结果表明,培养的滑膜细胞具有成纤维细胞的形态和特征,呈长梭形板向生长,RT-PCR方法检测波形蛋白表达较高.本研究成功地培养出了AA...     

成年鸡原代肝细胞的原位分离和长期培养

为探讨成年鸡原代肝细胞的分离条件和长期培养过程中的形态学特征,本试验采用改进胶原酶二步原位灌流法分离高活性鸡原代肝细胞,并优化培养液,得到肝细胞长期培养的初步条件.结果显示,分离得到的活性肝细胞达94%,培养3 d细胞活性最强;贴壁培养时可见到肝细胞较为明显的分化过程.     

DL-乙硫氨酸建立大鼠脂肪肝体外模型的研究

为研究建立大鼠脂肪肝体外模型的方法,试验采用原代肝细胞体外培养的方法培养大鼠肝细胞.对正常贴壁培养的肝细胞用1.0,2.0,3.0,4.0,5.0 mmol/L DL-乙硫氨酸处理,分别测定处理前及处理后24,48小时细胞培养基上清液中丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AsT)的活性以及细胞裂解液中甘油三酯(TG)的含量.结果表明:贴壁培养的正常肝细胞用DL-乙硫氨酸处理后,培养基中ALT、AST的活性均有不同程度升高,其中5.0 mmol/,L处理组升高明显,与对照组相比差异极显著(P<0.01);4.0,5.0 mmol/L组的细胞裂解液中TG含量与对照组相比差异极显著(P<0.01).结果说明试验建立了大鼠脂肪肝药物损伤的体外模型,可以为在细胞水平筛选治疗脂肪肝的药物提供参考模型.     

亚硒酸钠对游离脂肪酸致肝细胞凋亡和线粒体膜电位降低的保护作用

为研究亚硒酸钠对游离脂肪酸 (free fatty acid,FFA) 致体外培养大鼠肝细胞凋亡和线粒体膜电位降低的保护作用,本试验通过胶原酶两步灌流法分离培养肝细胞,在细胞培养液中添加FFA和亚硒酸钠 (0.1 μmol/L)测定细胞的凋亡情况、细胞功能和线粒体膜电位。结果表明,随着培养液中FFA浓度的升高,细胞凋亡加重,肝细胞功能受到影响,线粒体膜电位降低且FFA的这种作用呈剂量依赖性;亚硒酸钠能抑制由FFA引起的肝细胞的上述变化。     

大白鼠乳腺上皮细胞的体外培养与鉴定

 试验选取泌乳期大白鼠乳腺组织,采用机械剪切结合胰酶消化的方法,在体外进行乳腺上皮细胞原代分离培养及鉴定。结果成功培养出大鼠原代乳腺上皮细胞,经过角蛋白免疫组化鉴定染色后,呈现阳性反应。显微镜下观察,细胞呈鹅卵石样;经多次传代,细胞增殖仍然长势很好。     

Disease and destiny-mystery and mastery

When early people made their appearance, zoonotic infectious diseases were already waiting, but epidemic diseases did not appear in human history until people began to live in large numbers under conditions of close contact, mainly during the last 10,000 years. Disease has decimated urban populations, conquered armies, and disrupted society. The focus here is on (1) the plague of Athens and the Black Death; (2) smallpox, influenza, and rabies; (3) avian influenza prion diseases, and foot & mouth disease; and (4) emerging and re-emerging diseases. All have veterinary public health associations. In Athens, Greece, in 430 BC, when the Spartans ravaged the countryside, hordes crowded into Athens so that orderly movements, space in which to live, and adequate supplies of food became impossible. Crowding of any population fosters disease transmission; chaos and disorder enhance it all the more. Out of northern Egypt came a terrible plague from across the Mediterranean Sea. The identity of the plague of Athens remains unsure, but the well-considered conclusion is Rift Valley Fever, a mosquito borne, viral zoonosis. The Black Death, also called the Plague, raged in Asia for centuries. In 1347, the Black Death was brought by a ship out of Asia to Sicily. The scenes of devastation were repeated throughout Europe, with 90% or more of the people dying in city after city. Influenza, too, has been a cause of periodic human epidemics, but the great pandemic of influenza occurred in the last months of World War I. In the years of highest occurrence, more than half the world's population became clinically infected. If veterinary public health had been born earlier, it could have led to elucidating the epidemiology of influenza and the plagues of Athens, Europe, and Asia. In turn, smallpox had also caused continual tragedy. In 1796, Edward Jenner began to harvest pustules of cowpox from children or infected cows and inject them into susceptible children. In 1980, the World Health Organization declared that smallpox had been eliminated from the world. Rabies, though, still strikes terror. A number of animal diseases, broadly termed emerging and re-emerging diseases, need surveillance because they have the potential to impact human health. From late in 2003 to 2007, the highly pathogenic H5N1 influenza virus in poultry infected at least 121 people and caused 62 deaths in four countries. The prion diseases, too, all have very high numbers in concentrated contacts. To control these diseases, veterinary public health is essential, with diagnosis, epidemiological surveillance, clinical manifestations, and prevention as primary measures.     

Foot and mouth disease and bluetongue: differences and similarities

On 26th of july 2007 a new case of bluetongue was notified in the Netherlands and on 2nd of august 2007 foot-and-mouth disease was diagnosed in Surrey, England, which raised the threat of having both infections simultaniously in one area. Bluetongue and foot-and-mouth disease have a different pathogenesis, but symptoms may resemble each other at a later stage of infection. The pathogenesis and possible clinical symptoms of both infections are discussed and illustrated with pictures.     

德国荷兰畜禽产品质量安全交流与启示

2018年岁末,笔者有幸赴德国和荷兰,就畜产品质量安全控制及检测技术等进行了短期交流,收获颇丰,现具体介绍如下。一、交流情况德国面积35.73万平方千米,人口约8175.2万,是欧盟人口最多的国家,农业发达,机械化程度很高。德国的畜牧业以猪、牛、羊和禽类为主,畜牧业产值占农业生产总值的61%。德国虽然农业比重很小,但却是有机农业运动最早的发起国,也是目前世界上有机农产品生产与消费大国。     

奶牛肢蹄病及其防治

简介常见奶牛肢蹄病种类及其症状,从日粮营养、运动场地面结构、环境卫生、饲养管理、遗传育种、疾病管理等方面简析奶牛肢蹄病的病因,并提出相应的预防措施。     

畜禽物种多样性及其保护和利用

文章就我国畜禽物种多样性资源的现状、保护和利用等方面进行阐述,为生态脆弱的喀斯特地区畜牧业可持续发展提供参考。     

马的咽气癖及其防制措施研究进展

在动物的异常行为中以刻板行为最为常见,而咽气癖又是马最常见的口部刻板行为之一。作者就咽气癖在生理方面和心智方面对马体产生的影响、行为基础、诱因的研究进展及常见的防治措施进行综述。     

国内外蛋氨酸与赖氨酸市场分析与展望

近几年,世界饲用添加剂市场增长较快,预计今后将进一步增长,据世界粮农组织(FAO)首领会议的倡议,到2015年,将使世界营养不良的人口削减一半,其间,肉制品将会以每年2％的速度增长,特别是加快猪肉和禽肉增长。这将促进世界添加剂,尤其是蛋氨酸和赖氨酸需求量的增加。     

牛羊焦虫病诊断与防治技术

近几年来,辽宁省牛羊饲养业迅速发展,焦虫病的危害越来越明显,已成为危害牛羊的主要疾病之一。为了提高牛羊焦虫病的防治水平,保证我省牛羊饲养业的健康发展,笔者在参阅文献和同行们诊疗经验的基础上,结合自己的治疗实践,详细阐述了我省牛羊焦虫病近几年的流行特点和诊断防治方法。