牛皮肤成纤维细胞的体外培养与冻存   总被引：10，自引：0，他引：10  

任芳丽  李煜等 《中国牛业科学》2002,28(1):8-10

利用牛皮肤组织块直接培养法，得到牛皮肤细胞的原代培养物，再用酶消化法和反复贴壁法处理，能够纯化成纤维细胞。成纤维细胞的冻存是通过选用6种分别含有二甲基亚砜（DMSO）、甘油（GL）及乙二醇（EG）的保护液，以相同的冻前处理方法，对牛皮肤成纤维细胞进行缓慢冷冻，冰箱预冷平衡1－2h，逐步投入液氮（－196℃）中保存，再经37℃水浴解冻，Hanks液脱保护剂，以贴壁率评价冻存效果。结果表明，20％DMSO保护液对牛皮肤成纤维细胞表现出较好且稳定的冷冻保护效果，其平均贴壁率达87．9％。  相似文献   

成年(黄占)鹿耳皮肤成纤维细胞的分离与培养     

王启凤  王锋 《家畜生态》2005,(4)

本试验对1头黄占鹿的耳缘皮肤组织采用0.1%胶原酶(Ⅰ型)消化组织块法进行原代培养,反复传代纯化,获得了成纤维细胞,进行了生物学特性观察,并对各代细胞进行了常规冷冻保存。利用Photoshop软件对F12代细胞染色体进行了核型分析,结果表明该黄占鹿的染色体数目为68,常染色体中1号和7号为M染色体,其余的均为A常染色体;X染色体是最大的A染色体,Y染色体是最小的M染色体;该黄占鹿的核型式为4(M)+62(A),XY(A,M);对部分冷冻细胞进行了解冻培养,解冻存活率和贴壁率分别为61.04%、38.85%。  相似文献   

北京油鸡胚胎成纤维细胞系建立与生物学特性研究   总被引：12，自引：3，他引：12  

周向梅  马月辉  关伟军  文杰  李晗 《畜牧兽医学报》2005,36(3):209-215

采取组织块直接培养法，对北京油鸡胚胎组织进行原代和继代培养，成功地建立了成纤维细胞系。并对培养细胞进行了形态学、细胞生长动力学观察，以及核型和乳酸脱氢酶、苹果酸脱氢酶的同工酶分析。结果表明，该细胞系的群体倍增时间(PDT)为24h；细胞染色体中二倍体占主体，为76％～88％；乳酸脱氢酶、苹果酸脱氢酶同工酶电泳图谱与本室其它细胞系有明显差别；细菌、真菌、病毒、支原体检测呈阴性。该细胞系的建立，使北京油鸡这一国家重要种质资源在细胞水平上保存下来，也为基因组文库和体细胞克隆等研究提供了理想的生物材料。  相似文献   

白藜芦醇和褪黑素对绵羊胎儿成纤维细胞转染及卵母细胞体外成熟的影响     

姚昱君  李广栋  吴昊  马文奎  杨海  关盛宇  吕东颖  付瑶  朱天奇  姬鹏云  谭鑫星  赵万民  连正兴  张鲁  刘国世 《中国畜牧兽医》2021,48(12):4497-4507

试验旨在利用白藜芦醇(Re)和褪黑素(MT)提高绵羊胎儿成纤维细胞电穿孔转染(电转)效率、优化卵母细胞体外成熟体系,以提高转基因动物生产效率。将绵羊胎儿成纤维细胞分为6组:对照组、添加白藜芦醇高、中、低(R^-5、R^-6、R^-7)3种浓度组及白藜芦醇和褪黑素联合添加组(R^-5M^-5、R^-6M^-7),通过电转效率和细胞凋亡情况确定电转体系优化方案;通过对细胞周期、线粒体膜电位及活性氧(ROS)的检测研究其作用机制。将绵羊卵母细胞分为对照组及白藜芦醇和褪黑素(R^-5M^-5、R^-6M^-7)联合添加组,通过绵羊卵母细胞体外成熟后卵丘细胞扩展、第一极体排出及体外受精胚胎的卵裂率检测其对卵母细胞体外成熟的作用。研究结果显示,分离的绵羊胎儿成纤维细胞P3代增殖旺盛,细胞生长曲线呈典型的S型;与对照组相比,所有试验组胎儿成纤维细胞的电转效率均显著提高(P<0.05),R^-5M^-5组电转效率最高;R^-6M^-7组显著降低细胞凋亡率(P<0.05),R^-5、R^-6、R^-5M^-5和R^-6M^-7组均显著促进细胞增殖(P<0.05);R^-6、R^-7、R^-5M^-5和R^-6M^-7组G1期细胞增加,S期细胞相对减少,但差异均不显著(P>0.05);R^-6、R^-7、R^-5M^-5和R^-6M^-7组成纤维细胞内ROS水平均显著降低(P<0.05),R^-6、R^-5M^-5和R^-6M^-7组线粒体膜电位均显著增加(P<0.05);R^-6M^-7组显著促进绵羊卵丘细胞扩展、提高卵母细胞第一极体排出率和体外受精胚胎发育能力(P<0.05)。综上,白藜芦醇和褪黑素可以优化绵羊胎儿成纤维细胞电转体系,提高卵母细胞体外成熟效率,为白藜芦醇和褪黑素在转基因动物生产中的应用提供参考依据。  相似文献   

Effects of bFGF on brain edema,nerve function injury and autophagy related protein in rats with traumatic brain injury     

WANG Wen-hong  KONG Jun  LIN Xiao-xiang  LI Jian-xia  CHEN Lu-kui 《园艺学报》2019,35(2):320-325

AIM:To study the effects of basic fibroblast growth factor (bFGF) on brain edema, nerve function damage and autophagy related proteins in rats with head injury. METHODS:The rat model of craniocerebral injury (CI) was constructed. The rats were divided into control group, CI group, and low-, middle-and high-dose bFGF groups (n=10). The CI model was established in CI group, while the rats in control group were not given epidural impact. The rats in low-dose, middle-dose and high-dose bFGF groups were given bFGF at 2, 4 and 6 μg, respectively, by intraperitoneal injection after 30 min. The neurological function in the rats was evaluated by improved neurological function scoring. The rat brain tissues were taken, and the water content was detected. The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and IL-1β in the brain tissue were measured by ELISA. The malondialdehyde (MDA) content was analyzed by thiobarbituric acid method. The activity of superoxide dismutase (SOD) was examined by WST-8 assay. The glutathine peroxidase (GSH-Px) activity was detected by colorimetric method. The protein levels of autophagy related proteins LC3-Ⅱ and beclin-1 in the brain tissues were determined by Western blot. RESULTS:The neurological function score was increased significantly of the rats in CI group. The rat model of craniocerebral injury was successfully constructed. Neurological function scores in the rats in low-dose, middle-dose and high-dose bFGF groups were reduced, the water content of the brain tissue was also reduced (P<0.05). The levels of TNF-α, IL-6 and IL-1 β were decreased in the brain tissues (P<0.05), the content of MDA was declined (P<0.05), the activities of SOD and GSH-Px were increased (P<0.05), the protein levels of LC3-Ⅱ and beclin-1 were decreased, compared with the untreated rats in CI group (P<0.05). CONCLUSION:bFGF improves the nerve function of the rats with craniocerebral injury, reduces the water content of the brain tissue, reduces the expression of autophagic protein LC3-Ⅱ and beclin-1.The mechanism is related to the inhibition of inflammatory reaction and oxidative damage.  相似文献   

重组人骨形态发生蛋白2复合物对犬胫骨骨折愈合中4种相关细胞因子的作用     

梁倩  陈智华  亢心  年聚盆  王雯慧  韩鹏 《动物医学进展》2020,(1):56-60

研究重组人骨形态发生蛋白2(rhBMP-2)、碱性成纤维细胞生长因子(bFGF)、纤维蛋白凝胶(FG)与妥布霉素复合物加速骨折愈合。bFGF与rhBMP-2和妥布霉素以纤维蛋白胶为载体复合,将具有抗生素缓释系统的复合物注入犬胫骨骨折处,并做内固定,于术后第2周,采集犬胫骨骨折处样品,每隔2周采集1次,共采集4次,将每次采集的样品制成组织切片,采用免疫组织化学的方法检测4种因子,并对组织切片进行分析,用多功能真彩色细胞图象分析管理系统,分析所得数据用SPSS18.0版统计软件进行统计学分析。血管内皮细胞生长因子(VEGF)在试验组第2周表达强阳性,而对照组阳性表达较弱;前4周,试验组血小板源性生长因子(PDGF)阳性率呈上升趋势,强于对照组;前8周,试验组胰岛素样生长因子(IGF)和转化生长因子-β1(TGF-β1)阳性率均高于对照组。说明bFGF与rhBMP-2和妥布霉素以纤维蛋白胶为载体的复合物在骨折愈合具有中促进细胞增殖、黏附、趋化、分化,以及骨折末端血管生长和形成的作用。  相似文献   

Study on Antiviral Effect of Polysaccharides from Lagenaria siceraria (Molina) Standl.on NDV in vitro     

ADELIJIANG Wusiman  DELIBAIER  NUERAILI Maitiniyazi  BUHELIQIANMU Yiming  SAIFUDING Abula 《中国畜牧兽医》2016,43(7):1869-1875

To research on polysaccharides from Lagenaria siceraria (Molina) Standl.against Newcastle disease virus (NDV).Total polysaccharide and four different solvent extractions of Lagenaria siceraria (Molina) Standl.were extracted by the methods of hot water extraction and ethanol precipition in this study.The content of polysaccharide was measured by phenol-sulfuric acid method and infrared spectroscopy method.The safe concentration and growth of chick embryo fibroblast (CEF) were assayed by MTT method,in order to facilitated the comparison under the same level,the safe concentration was united as 78.125 μg/mL.Under the safety range of concentration,detected the block-virus activity,anti-virus activity and virus-killing activity of polysaccharides through the ways of pre-adding polysaccharides,post-adding polysaccharides and adding polysaccharides with NDV.The results showed that the direct inactivation and propagation inhibition activity of total polysaccharide and four different solvent extractions were stronger than anti-absorption function.Anti-virus inhibition rate of 70%,80% gradient alcohol precipitation of polysaccharides from Lagenaria siceraria (Molina) Standl.(LSP₇₀,LSP₈₀) and total polysaccharide (LSP_t)were 40.41%,44.54% and 61.85%,virus-killing inhibition rate were 44.74%,58.76% and 59.38%.LSP₈₀ had the highest virus inhibition rate as 37.14% in the block-virus activity of those five polysaccharides.In summary,70%,80% gradient alcohol precipitation and total polysaccharide in polysaccharides possessed better activity and would be as the materials for further research.  相似文献   

Expression of fibroblast growth factor 23 by canine soft tissue sarcomas          下载免费PDF全文

M. R. Hardcastle  K. E. Dittmer 《Veterinary and comparative oncology》2016,14(3):306-317

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siRNA干涉禽流感病毒PA基因在鸡胚成纤维细胞中的表达     

马莉  朱金凤  汤承  李明义  李定霏  岳华 《河南农业大学学报》2007,41(4):438-441

用阳离子脂体质转染试剂将人工合成的小干扰RNA(Small interference RNA,siRNA)转染鸡胚成纤维细胞(CEF).结果表明,针对禽流感病毒(AIV)H5N1亚型核蛋白(NP)基因的siRNA能在AIV感染后1,2,3 h显著抑制NP基因的表达,表达水平分别降低3,1.4和2.5倍.  相似文献   

鲁西黄牛耳缘组织成纤维细胞的体外培养   总被引：1，自引：0，他引：1  

郭瑜  关伟军  赖松家 《畜禽业》2008,(4):12-13

实验以鲁西黄牛耳缘组织为研究材料,采用组织块贴壁培养法对其进行了成纤维细胞的体外培养。通过细胞的传代培养、形态学和动力学观察与分析,结果表明,所建立的鲁西黄牛耳缘组织成纤维细胞系具有典型的成纤维细胞形态,胞体均为梭形和不规则三角形,且细胞丰满;细胞核形态为卵圆形,并位于胞质中央,核仁清晰。在细胞生长于繁殖过程中亦经历了潜伏期、指数增生、停滞期和衰退死亡期等4个时期。细胞群体倍增时期为72h。实验结果表明,已成功地培养了鲁西黄牛耳缘组织成纤维细胞。  相似文献