共查询到19条相似文献，搜索用时 906 毫秒
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传代细胞株和传代细胞系在超低温冷冻下,细胞既可长期保存,又能随时取用,对随时随地有检疫任务的检疫单位,特别适用.细胞冷冻保存比细胞连继传代培养能节省时间与材料,减少由于多次传代引起的遗传变异,还可防止在长期连续传代过程中可能发生的细菌或支原体污染. 相似文献
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对不同鲜度的猪肉、牛肉、羊肉、鸡肉和鱼肉进行活菌总数、革兰氏阴性菌数、大肠菌群数及挥发性直基氮（TVB－N）测定，并与LAL试验（鲎变形细胞溶解物试验）进行平行对比研究，证实肉品的鲜度与其细菌总数、革兰氏阴性菌数及LAL指数呈负相关性。应用LAL试验可在短时间（1－2小时）内快速判定肉品的细菌污染程度及鲜度，比传统的细菌培养计数法大大缩短了时间，适合现场大批量肉品细菌污染程度及鲜度的快速检测。 相似文献
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我国科学家经过长期的努力,成功地研制出了猪瘟活疫苗(传代细胞源),即一般所称的"猪瘟传代细胞苗",简称猪瘟ST苗.猪瘟传代细胞苗是一种优质高效的新型疫苗,它具有高度不易污染外源病毒而特别安全、高效、无应激、价格合理等特点.研究人员预测猪瘟传代苗将在未来猪瘟的预防控制方面起重要作用. 相似文献
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为生产检验需要，将生长好的ＢＴ细胞分别放在３７℃、２５℃、１５℃的温箱中，发现在２５℃下，细胞维持到需传传代的最长天数为５０－６０ｄ，传代前将ＢＴ细胞在３７℃预热１６－２４ｈ菌传代，即可获得生长良好的传代细胞。 相似文献
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对霉形体进入DK传代细胞内,并进行增殖的情况,作了详细的电镜观察.结果如下:(1)霉形体靠近传代细胞膜表面呈串珠样排列;霉形体与传代细胞膜表面接触部位融合增厚,电子密度增高,向内凹陷,将霉形体裹入细胞质,形成由膜结构包着的包涵体.(2)在传代细胞质中,除可见由膜结构包裹着的霉形体性包涵体外,还可见没有任何膜包着的散在的霉形体,并且均可见到霉形体的裂殖增殖相和出芽增殖相.(3)在传代细胞核的核周池和核质中,均见到典型的霉形体及其裂殖增殖相.(4)霉形体在传代细胞中大量的增殖,致使细胞崩解,霉形体即被释放出来.(5)霉形体使传代细胞出现严重的超微病变.本文还讨论了传代细胞培养污染霉形体后难以救治的根本原因是,霉形体在传代细胞内增殖,故仅仅着眼于消除培养液中的霉形体是不能达到预期结果的. 相似文献
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兔出血症病毒较难适应细胞,病毒在细胞上传代以后不仅血凝价下降较快,并对健康兔的致病性丧失也较快。研究该病毒的细胞传代材料刺激健康兔产生免疫力的能力,可进而探讨应用细胞材料制备疫苗的可能性。本文将介绍我们用细胞上传代的病毒 相似文献
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Effects of Serum Deprivation and Cycloheximide on Cell Cycle of Low and High Passage Porcine Fetal Fibroblasts 总被引：1，自引：0，他引：1
MD Goissis HVA Caetano MG Marques FRO de Barros WB Feitosa MP Milazzotto M Binelli MEOA Assumpção JA Visintin 《Reproduction in domestic animals》2007,42(6):660-663
Arrest of cells in G0/G1 cell cycle phase is desired for nuclear transfer procedures. Serum starvation and cell cycle inhibitors are different ways to induce synchronization of the cell cycle. This study investigated the effects of serum starvation and cycloheximide (CHX) on the cell cycle of low (5th) and high (15th) passages fetal porcine fibroblasts. Cell cycle phases were determined using fluorescent activated cell sorting. Fifth passage fibroblast cultures had higher (p < 0.05) proportion of cells in G0/G1 only after 72 h of serum starvation (77.60 ± 0.65) when compared with non‐starved cells (71.44 ± 1.88). Serum starvation for all periods tested induced an increase (p < 0.05) on proportion of cells in G0/G1 on the 15th passage. No significant differences were observed on the 5th passage cultures exposed to CHX, although, on the 15th passage an increase on proportion of cells was observed after all periods of exposure (p < 0.05). These data indicates that high passage cells in vitro are more susceptible to serum starvation and CHX G0/G1 synchronization. 相似文献
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M.O. Asagba Y.K. Ssentongo R.H. Johnson J.R. Smith 《Veterinary immunology and immunopathology》1981,2(1):87-94
A method is described by which cell lines can be readily developed from bovine peripheral leucocytes. Fifteen cell lines have been developed from 25 attempts, passage levels up to 60 being reached. The cell lines are aneuploid and predominantly epithelial, show split ratio capabilities of 1:4 to give monolayers with 5 days of routine passage, and have high resistance to laboratory contamination with bacterial and fungal agents. Data are given concerning establishment, morphology, viral susceptibility and chromosomal counts of established cell lines. 相似文献
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《Journal of aquatic animal health》2013,25(4):365-372
Abstract Largemouth bass virus (LMBV), a recently discovered iridovirus found in the eastern United States, is usually detected by isolation in cell culture. Although LMBV will replicate in several cell lines, optimal cell culture methods for the detection of this virus have not been determined. We tested inoculation method, adsorption time, incubation temperature, and various cell lines to determine the conditions that would provide the most sensitive cell culture assay for LMBV. The optimal inoculation procedure tested was to remove the culture medium from the culture well before the addition of the inoculum, and the optimal adsorption procedure tested was to allow the virus to adsorb for 40 min while the plates were on an orbital shaker. Following inoculation, incubation at 30°C resulted in a higher number of viral plaques than incubation at 25°C or 32°C. Four cell lines (bluegill fry (BF-2), fathead minnow (FHM), epithelioma papulosum cyprini, and channel catfish ovary cells) inoculated with LMBV had similar susceptibility to infection. Similar percentages of LMBV-positive samples were detected in BF-2 and FHM cell cultures inoculated with homogenized organ samples from largemouth bass Micropterus salmoides; however, the use of two cell lines increased the number of infected samples discovered. A blind passage also increased the number of positive samples detected in cell culture. Subcultivation to confirm virus-positive samples was useful for reducing false-positive results. 相似文献
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Virus Pneumonia of Pigs; Attempts at Propagation of the Causative Agent in Cell Cultures and Chicken Embryos 总被引：1，自引：1，他引：0 
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下载免费PDF全文 Two strains of the agent of virus pneumonia, were tested for the ability to propagate in 12 types of cell cultures and in chicken embryos. The 5 primary cell cultures used were: swine kidney, lung, bone marrow, testicle, and chicken embryo kidney; and the 7 serial passage cell cultures were: swine kidney, kidney-tumor, testicle, bone-marrow, bovine kidney, and human cervical carcinoma (HeLa). The agent of virus pneumonia was propagated in primary swine kidney and in HeLa cell cultures as shown by the production of typical gross and microscopic lesions in pigs inoculated with cell future fluids. Third passage cell culture fluids, produced typical gross lesions in pigs, but fourth passage cell culture fluids produced only microscopic lesions, and no lesions were produced by sixth and eleventh passage fluids. Control pigs receiving fluids from uninoculated cell cultures remained free of gross or microscopic lesions, as did uninoculated controls. Cytopathic effects were not detected in any of the inoculated cell cultures and no cellular changes were detected by staining with Giemsa stain or acridine orange.
Neither lesions nor deaths occurred in chicken embryos inoculated with both strains of virus pneumonia virus. Pneumonia was not produced in pigs inoculated with suspensions from second chicken embryo passage of the 2 strains inoculated by the chorioallantioic sac, the amniotic sac, and the yolk sac routes.
Identical gross and microscopic lesions were produced in pigs inoculated with either pneumonic lung suspensions or with virulent cell culture fluids. Gross lesions consisted of areas of light to reddish-purple consolidation usually limited to the anterior, cardiac, and intermediate lobes of the lungs. Pleuritis and pericarditis were never present in experimentally produced virus pneumonia. The microscopic lesions were characterized by: 1. perivascular and peribronchiolar lymphoid infiltration and hyperplasia, 2. alveolar interstitial thickening and infiltration, and 3. alveolar exudates consisting of alveolar cells, lymphocytes, plasma cells, and neutrophiles.
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Hasebe R Kimura T Nakamura K Okazaki K Ochiai K Wada R Umemura T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2002,64(10):907-912
Intracerebral inoculation of field-isolates as well as established strains of equine herpesvirus-1 (EHV-1) in suckling mice results in viral replication in neurons and glial cells and induces encephalitis. By intraperitoneal (i.p.) inoculation, no histological lesion was observed in the central nervous system (CNS) in suckling mice with the EHV-1 HH1 strain (HH1), whereas a neuroadapted variant (NHH1) produced by serial passage of HH1 in the mouse brain caused severe encephalomyelitis after i.p. inoculation. The purpose of this study was to determine the route of neuroinvasion after i.p. inoculation of NHH1 and to clarify the effects of the brain passage on viral neuroinvasion. NHH1, but not HH1, targeted splenic and pulmonary macrophages and omental fat cells on days 1 and 2 post-inoculation (p.i.). From days 1 to 3 p.i., cell-associated viremia was occurred in NHH1-infected mice, but not in HH1-infected mice. On day 4 p.i., viral antigen was detected in a few endothelial cells, perivascular glial cells and neurons in the CNS in NHH1-infected mice. The number of viral antigen-positive cells increased markedly after day 5 p.i. In contrast, no viral antigen-positive cell was detected in the CNS in HH1-infected mice, except for a few nerve cells in the thoracic cord on day 4 p.i. These results suggest that NHH1 neuroinvasion is hematogenous and is correlated with enhanced extraneural virus growth. 相似文献
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为了解市售鲜猪肉葡萄球菌和大肠杆菌污染情况,本试验从贵州省9个地区农贸市场采集鲜猪肉样本106份,3个地区屠宰场运输车辆采集棉拭子样本36份,采用传统生化法鉴定2种优势菌落;PCR法快速检测葡萄球菌和大肠杆菌。结果显示,鲜猪肉和棉拭子样本葡萄球菌的分离率为23．5％（38／162）和31．9％（23／72）,大肠杆菌的分离率为15．4％（25／162）和13．9％（10／72）;2种细菌可扩增出目的条带。结果表明,市售鲜猪肉及运输车辆葡萄球菌和大肠杆菌污染严重,本试验成功运用PCR法对这2种病原菌进行快速检测。 相似文献
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滩羊成纤维细胞系的建立及其生物学特性研究 总被引：2，自引：1，他引：1
试验以滩羊耳缘组织为材料,采用组织块贴壁培养法和细胞冷冻保存技术构建了35个滩羊成纤维细胞系,并对其进行了形态学、生长动力学、细胞活力测定、核型分析、微生物检测和荧光蛋白报告质粒(PEGFP-N3)基因转染等生物学特性的研究。结果表明,细胞群体倍增时间(PDT)约为36h,细胞冻存后活力为95.8%,细胞染色体中二倍体(2n=54)占主体,镜检细胞的百分比约为97.5%。细菌、真菌、病毒和支原体检测结果为阴性,该细胞库的各项指标均达到ATCC细胞系鉴定标准。 相似文献
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为验证猪链球菌2型荧光PCR检测方法对临床样品检测的敏感性和适用性以及该方法所拥有的独特优点,分别用荧光PCR法、常规PCR法和细菌分离法对人工感染猪链球菌2型的小鼠肝脏和发病猪的心、肝、脾、肾等实质器官和血液、喉拭子进行抗原检测。结果显示,荧光PCR法检出率为70.8%,明显高于普通PCR法（检出率为20.8%）,也高于常规细菌分离法（检出率为45.8%）。由于临床样品常常会被其他细菌污染,细菌分离法很难准确分离到链球菌。但荧光PCR法不受其他细菌污染的影响,对实验室培养的猪链球菌2型菌液,该方法检测滴度可达10-6/0.1mL（42～52 CFU/0.1 mL）,而普通PCR方法检测滴度仅为10-4。 相似文献
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贵德黑裘皮羊耳成纤维细胞系的建立和生物学特性的研究 总被引：4，自引：1，他引：4
以贵德黑裘皮羊耳缘组织为材料，采用组织块贴壁培养法和细胞冷冻技术成功构建了成纤维细胞系，并对培养细胞进行了形态学、生长动力学观察、细胞活力测定、核型分析和微生物检测。结果表明：细胞群体倍增时间（PDT）约为36h，冻存前细胞活力为96．2％，以DMEM／F-12＋20％FBS中添加10％DMSO冻存成纤维细胞，解冻后细胞活力为93．6％，24h后的贴壁率为85％。在传代10次之前，细胞染色体中二倍体（2n=54）占主体，约为82％～90％，细菌、真菌、病毒、支原体检测为阴性。该细胞系的建立，使贵德黑裘皮羊这一国家重要种质资源在细胞水平上保存下来，也为体细胞克隆等研究提供了理想的生物材料。 相似文献