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1.
圆斑星鲽连续性鳃细胞系的建立 总被引：1，自引：1，他引：0
为建立圆斑星鲽(Verasper variegatus)鳃细胞系,本文采用胰蛋白酶消化法启动了圆斑星鲽鳃组织的体外培养,并通过对培养液配方和培养条件的优化成功地进行了圆斑星鲽鳃细胞的原代培养和继代培养。结果显示,圆斑星鲽鳃细胞的最适培养液为含有20%胎牛血清(FBS)的DMEM/F-12培养液,培养液最适pH值为7.2,最适培养温度为22℃;通过向20%FBS-DMEM/F-12培养液(pH=7.2)中添加N-乙酰葡萄糖盐酸盐、羧甲基壳寡糖、碱性成纤维细胞生长因子(bFGF)及I型胰岛素样生长因子(IGF-I),在22℃成功启动了圆斑星鲽鳃细胞的原代培养,细胞的贴壁、生长和分裂状态良好,为典型的成纤维样细胞形态,30 d即可形成汇合的细胞单层;经连续继代培养后,已成功建立了连续性圆斑星鲽鳃细胞系,现已继代培养至第76代;该细胞系细胞经液氮冻存复苏后仍保持其原有的成纤维样细胞形态,其贴壁生长状态和增殖速度也与冻存前无差异;生长特性鉴定结果显示,第60代圆斑星鲽鳃细胞系细胞的群体倍增时间为43.6 h,表明细胞的生长分裂依然十分旺盛。染色体分析结果显示,圆斑星鲽鳃细胞系细胞虽然出现了染色体的非整倍性,但其特征性染色体数目仍为46条,并具有其典型的二倍体核型特征(46 t),表明所建立的细胞系确为圆斑星鲽鳃细胞系。该细胞系的建立对于病毒与宿主相互作用机理的研究及病毒疫苗的开发具有重要的理论和应用价值。 相似文献
2.
褐点石斑鱼三种组织细胞系的建立 总被引：2，自引：1，他引：1
为建立褐点石斑鱼鳍、心脏和鳔组织细胞系,本文利用不同培养液和培养温度对褐点石斑鱼鳍、心脏和鳔组织细胞进行了原代培养和传代培养。实验结果显示,在24℃培养于含有羧甲基壳寡糖、碱性成纤维样生长因子、I型胰岛素样生长因子及20%胎牛血清的DMEM/F12培养液(pH=7.2)中的3种细胞,均为成纤维样细胞,其生长分裂状态最佳,可以持续稳定传代。第60代褐点石斑鱼鳍、心脏和鳔细胞的群体倍增时间分别为50.6 h、40.3 h和43.3 h,其特征性染色体数目均为48条。目前鳍、心脏和鳔细胞已分别传至第90代、第70代和第75代,已成功建立了3种细胞的连续性细胞系,为鱼类病毒与宿主细胞相互作用机制等鱼类病毒学基础研究,以及鱼类病毒的分离、鉴定、繁殖及病毒疫苗研制等奠定了基础。 相似文献
3.
大黄鱼鳍细胞系的建立及久效磷对其毒性作用的研究 总被引：1，自引：1，他引：0
为了建市大黄鱼(Pseudosciaena crocea)鳍细胞系,为其细胞毒理学、病毒学与细胞工程等研究奠定基础,本文采用酶消化法使用含有20%胎牛血清(FBS)的DMEM/F12、Leibovitz L-15和DMEM培养液(pH7.2)在22～28℃分别启动了大黄鱼鳍组织的体外培养,通过添加促细胞贴壁和分裂的物质在最适堵养条件下对大黄鱼鳍细胞进行了原代培养和继代培养,并研究了久效磷对大黄鱼细胞系鳍细胞的毒性作用.体外培养结果显示,大黄鱼鳍细胞的最适培养液为20%FBS-DMEM/F12,最适培养温度为25℃,体外培养鳍细胞的形态主要为成纤维细胞样,22 d后便可形成汇合细胞单层,经过连续继代培养成功建立了大黄鱼的连续性鳍细胞系,目前已传至第112代;对第60代大黄鱼鳍细胞系细胞的鉴定结果显示,其群体倍增时间为50.96 h,分裂状态十分旺盛,虽然出现了染色体的非整倍性,但其特征性染色体数目仍为48条,并具有正常的6m+6sm+36t二倍体核型,证明所建立的细胞系确为大黄鱼鳍细胞系.不同浓度久效磷处理的检测结果显示,大黄鱼鳍细胞对久效磷敏感,20～160 μg/mL久效磷可引起鳍细胞乙酰胆碱酯酶活性的持续性显著降低,引起超氧化物歧化酶和谷胱甘肽-S-转移酶活性的显著升高并随着浓度的升高而逐渐降低,久效磷对该细胞系细胞的48 h半抑制浓度(IC50)为43.05 μg/mL,,证实久效磷对大黄鱼鳍细胞具有显著的毒性作用. 相似文献
4.
为建立来源于锦鲤的细胞系,本文采用组织块法,对来源于锦鲤(Cyprinus carpio)鳍条、吻端、肌肉、心脏、鳔、肠道、卵巢等组织的细胞进行原代培养和传代培养.在25℃下,添加20％胎牛血清、0.2μg/mL表皮生长因子(EGF)和25ng/mL成纤维生长因子(FGF)的L-15培养基中进行培养.结果显示,鳍条、心脏、鳔、吻端、肌肉、肠道、卵巢分别在原代培养第3、6、12、13、15、3、3天,有细胞从组织块迁出;对长至单层的鳍条、心脏、肌肉细胞进行传代培养,鳍条细胞已传至第39代,心脏和肌肉细胞分别传到第2代和第4代;鳍条、心脏和肌肉细胞分别呈现上皮细胞样、上皮细胞样和上皮细胞样与成纤维状混合型.第6代锦鲤鳍细胞的染色体计数,结果显示,细胞染色体数目分布范围为55～154条,2n=100.病毒敏感性实验,发现锦鲤鳍条细胞对草鱼出血病病毒(GCRV)和鲤春病毒血症病毒(SVCV)都敏感,且在24h内出现细胞病变(CPE);但对斑点叉尾鮰病毒(CCV)不敏感.锦鲤鳍条细胞系的建立为后期建立更多的锦鲤细胞系和鱼类病毒研究奠定了基础. 相似文献
5.
宽纹虎鲨软骨细胞体外培养的启动 总被引：5，自引：1，他引：4
以活体宽纹虎鲨(Heterodortus japonicus)的鳃软骨和鳍软骨组织为材料,利用透明质酸酶(hyaluronidase)、Ⅱ型胶原酶(type Ⅱ collagenase)和胰蛋白酶(trypsin)进行消化获得游离软骨细胞,对所得游离细胞连同软骨组织分别用含20％小牛血清的MEM培养液进行体外培养。对软骨细胞体外培养条件的优化结果显示,宽纹虎鲨鳍软骨细胞体外培养的最适pH为7．2～7．6、最佳温度为24℃,成纤维细胞生长因子(bFGF,basic fibroblast growth factor)对软骨细胞作用不显著。培养于pH7．2～7．6的MEM培养液(含20％小牛血清)中的宽纹虎鲨鳍软骨组织,在24℃启动培养3d后便有软骨细胞开始陆续从组织中迁出,新迁出细胞的形态多为圆形,随着培养时间的延长,部分细胞形态逐渐转变成上皮样。 相似文献
6.
探讨大菱鲆病毒疫苗的研制,以牙鲆鳃细胞(FG细胞)为增殖体系,利用细胞病变效应(CPE)为判断指标,对大菱鲆红体病虹彩病毒(Turbot reddish body iridovirus,TRBIV)的体外最适增殖条件等进行了研究。结果表明,病毒接种量、病毒吸附时间、细胞培养条件对TRBIV的体外增殖均有明显的影响,以0.2 mL病毒液接种FG细胞、吸附2 h、用含10%胎牛血清的MEM培养液于22℃培养时,病毒的增殖速度最快。研究还发现,病毒的收获时间和病毒液冻融次数对所得病毒的感染力均有显著影响,在最佳增殖条件下,病毒在FG细胞中增殖至第4天时进行收获,且只冻融1次,其感染活性最强,反复冻融后病毒的感染活性会显著降低。 相似文献
7.
利用牙鲆鳃细胞系分离和培养淋巴囊肿病毒 总被引：1，自引：4，他引：1
本文利用牙鲆鳃细胞系进行了养殖牙鲆淋巴囊肿病毒的分离及培养 ,并通过电镜对培养细胞中淋巴囊肿病毒的形态及感染循环进行了初步研究。将病鱼的淋巴囊肿组织无菌滤液接种牙鲆细胞系 ,细胞出现了明显的细胞病变 ( Cytopathic effect,CPE)。电镜观察在培养细胞的胞质中有病毒的包涵体 ,胞质中散在 6角形、5角形或圆形的病毒粒子 ,大小为 10 0～ 140 nm之间。在感染细胞的线粒体中也存在大量的病毒颗粒。 相似文献
8.
凡纳滨对虾血淋巴、类淋巴细胞培养 总被引：8，自引：0，他引：8
以 ×L-15 培养基为基础,添加不同比例的胎牛血清和对虾肌肉提取液配制成不同的培养液,当 22×L-15培养基、胎牛血清和对虾肌肉提取液的比例分别为 60 %、20 %、20 %时,培养效果较好。培养液 pH 维持在7.0 ~ 7.2时血淋巴和类淋巴细胞生长最好,且类淋巴细胞迁出迅速。绝大部分血淋巴细胞贴壁迅速并伸展成多角样或披针形,一般存活 7 ~ 9 d,少部分呈悬浮状态的血淋巴细胞存活达 60 d 以上。培养中如未及时去除对虾血清,血淋巴细胞迅速衰老并死亡。类淋巴组织细胞 6 d可形成单层,迁出细胞主要有两种形态: 多角样和成纤维样,细胞可存活 16 d以上。 相似文献
9.
为了探明姬松茸提取物的体外抗病毒感染活性及其作用方式,进而为水产养殖鱼类高效抗淋巴囊肿病毒(lymphocystis disease virus,LCDV)感染活性物质的开发和鲆蝶类淋巴囊肿病的防治奠定基础,利用热水浸提和酒精沉淀法得到了5种姬松茸提取物组分(E1～5),并利用MTT、细胞病变程度观察等方法研究了5种组分对LCDV感染体外培养大菱鲆鳍细胞(turbot fin cells,TF细胞)的影响作用.细胞毒性实验结果显示,5种组分对体外培养TF细胞均无毒性.细胞病变程度结果表明,本文所得5种姬松茸提取物组分尤其是E5组分具有显著的抗LCDV感染TF细胞的活性.不同方式感染的实验结果进一步显示,5种姬松茸提取物组分抗LCDV感染TF细胞的作用可能主要是通过直接灭活病毒和/或阻断病毒吸附细胞来实现的. 相似文献
10.
云纹石斑针淋巴囊肿病病变过程的超微研究 总被引：3，自引：1，他引：3
于１９８９１－９９０年，对海水网箱养殖的云纹石斑鱼自然感染和人工感染的淋巴囊肿病进行电镜观察。观察结果表明，自然感染和人工感染的淋巴囊肿细胞之间并无区别，经们均含有许多包涵体和病毒颗粒；主要描述用患淋巴囊肿病鱼的皮肤结节匀浆对实验鱼 下注射而感染的淋巴囊肿病的超微病理过程，包括病毒颗粒在成纤维细胞胞膜上的附着和成纤维细胞及其内含物的形态变化等过程；此外，病鱼心脏、肝脏、脾脏、肾脏和肠上皮也有不同程 相似文献
11.
Derek V. Ellis 《Marine Georesources & Geotechnology》2001,19(1):51-63
This article reviews information recently available from existing marine and coastal mining for responses to environmental issues affecting marine mining at different depths. It is particularly but not exclusively concerned with those issues affecting seabed biodiversity impact and recovery. Much information has been gathered in the past 10 years from shallow mining operations for construction aggregate, diamonds, and gold, from coastal mines discharging tailings to shallow and deep water, and from experimental deep mining tests. The responses to issues identified are summarized in a series of eight tables intended to facilitate site-specific consideration. Since impacts can spread widely in the surface mixing layer SML, and can affect the biologically productive euphotic zone, the main issues considered arise from the depth of mining relative to the SML of the sea. Where mining is below the SML, the issue is whether it is environmentally better to bring the extraction products to the surface vessel for processing (and waste discharge), or to process the extraction products as much as possible on the seabed. Responses to the issues need to be sitespecific, and dependent on adequate preoperational environmental impact and recovery prediction. For deep tailings disposal from a surface vessel, there are four important environmental unknowns: (1) the possible growth of "marine snow" (bacterial flocs) utilizing the enormous quantities of fine tailings particles (hundreds or thousands of metric tons per day) as nuclei for growth, (2) the possibility that local keystone plankton and nekton species may migrate diurnally down to and beyond the depth of deep discharge and hence be subjected to tailings impact at depth, (3) the burrow-up capability of deep benthos and their ability to survive high rates of tailings deposition, and (4) the pattern and rate of dispersion of a tailings density current through the deep water column from discharge point to seabed. Actions to obtain relevant information in general and site-specifically are suggested. 相似文献
12.
G. Parthiban 《Marine Georesources & Geotechnology》2000,18(3):223-235
Particle fluxes were measured 7 m above the sea bottom during the predisturbance, disturbance, and postdisturbance periods by using time series sediment traps attached to seven deep-sea moorings deployed in the INDEX experiment site in the Central Indian Basin. The predisturbance particle fluxes varied between 22.3 to 55.1 mg m -2 day -1 . Increased and variable particle fluxes were recorded by the sediment traps during the disturbance period. The increase observed was 0.5 to 4 times more than the background predisturbance fluxes. The increases in particle fluxes (~4 times) recorded by the sediment trap located in the southwestern direction (DMS-1) were the greatest, which could be the result of preferential movement of resuspended particles generated during the deep-sea benthic disturbance along the general current direction prevailing in this area during the experimental period. Also, the traps located closer to the disturbance area recorded greater fluxes than did the traps far away, across the Deep Sea Sediment Resuspension System path. This variability in recorded particle fluxes by the traps around the disturbance area clearly indicates that physical characteristics such as grain size and density of the resuspended particles produced during the disturbance had an important effect on particle movement. The postdisturbance measurements during ~5 days showed a reduction in particle fluxes of ~50%, indicating rapid particle settlement. 相似文献
13.
Arne Schnack-Friedrichsen Angela M. Davis Jim D. Bennell Dei G. Huws 《Marine Georesources & Geotechnology》2001,19(4):221-243
The interdependence between the seismo-acoustic properties of a marine sediment and its geotechnical/physical parameters has been known for many years, and it has been postulated that this should allow the extraction of geotechnical information from seismic data. Though in the literature many correlations have been published for the surficial layer, there is a lack of information for greater sediment depths. In this article, a desktop study on a synthetic seafloor model illustrates how the application of published near-surface prediction equations to subsurface sediments (up to several tens of meters burial depth) can lead to spurious predictions. To test this further, acoustic and geotechnical properties were measured on a number of sediment core samples, some of which were subjected to loading in acoustically-equipped consolidation cells (oedometers) to simulate greater burial depth conditions. For low effective pressures (representing small burial depths extending to around 10 meters subsurface), the general applicability of established relationships was confirmed: the prediction of porosity, bulk density, and mean grain size from acoustic velocity and impedance appears generally possible for the investigated sedimentary environments. As effective pressure increases through, the observed relationships deviate more and more from the established ones for the near-surface area. For the samples tested in this study, in some instances increasing pressure even resulted in decreasing velocities. There are several possible explanations for this abnormal behavior, including the presence of gas, overconsolidation, or bimodal grain size distribution. The results indicate that an appropriate depth correction must be introduced into the published prediction equations in order to obtain reliable estimates of physical sediment properties for greater subsurface depths. 相似文献
14.
海上大直径钢管桩打桩过程中,桩周土体受到强烈扰动而发生强度弱化,掌握桩周土体强度弱化规律对于准确预测打桩过程、保证工程安全具有重要意义。为研究土体强度弱化规律,开展了环剪试验模拟打桩对桩周土体的扰动,测试土体强度随剪切速率的变化规律,建立了描述土体强度弱化规律的拟合公式,引入到打桩分析软件中。研究结果表明：土体的强度折减程度不仅与土体本身的性质有关还受到土体的埋深和剪切速率的影响,埋深越深土体强度折减程度越低,剪切速率越高土体强度折减越高,在打桩分析中可采用这里推荐的线性折减方法来模拟不同深度处土体强度的折减规律。 相似文献
15.
The advanced piston corer (APC) has been used by the Ocean Drilling Program since 1985 for recovering soft sediments from the ocean floor. The pullout force measured on extracting the core barrel from the sediment is shown to correlate with the average shear strength of the sediment core measured in the ship's laboratory. A simple rule of thumb is derived relating the shear strength of the sediment to the pullout force. Multiple APC holes at individual sites allow the consistency of the pullout measurements to be assessed. The effects of different operational procedures during APC coring are also explored. Although generally applicable, the correlation between pullout force and laboratory measurements of shear strength breaks down for some APC holes, possibly because of the disturbance of some sediment types during the APC coring process. A better understanding of the physical process of APC coring, and its effect on the properties of the sediment both inside and immediately outside the core barrel, would indicate what confidence can be put on the measurement of pullout force as a way of evaluating the in situ shear strength of deep sea sediments. 相似文献
16.
Shigehisa Nakamura 《Marine Geodesy》2000,23(1):55-61
Specific properties of the interannual sea level variations and annual tides in the Northwestern Pacific were studied. Several tide stations were monitored. The monthly mean sea level for the year of 1995 was analyzed at each tide station. A seismic event in 1995, some tectonic activity around the subject area, and the Kuroshio (the oceanic western boundary current) may possibly contaminate results which would have occurred from the astronomical annual tide alone. 相似文献
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《海洋学报(英文版)》2014,(11):161-F0003
<正>Acta Oceanologica Sinica(AOS)is a comprehensive academic journal edited by the Editorial Committee of Acta Oceanologica Sinica and is designed to provide a forum for important research papers of the marine scientific community which reflect the information on a worldwide basis.The journal publishes scholarly papers on marine science and technology,including physics,chemistry,biology, 相似文献