转pGH基因猪与非转基因猪成纤维细胞系的建立及鉴定

发布时间：2018-03-21 05:22

本文选题：pGH基因　切入点：转基因猪　出处：《中国兽医杂志》2017年06期 　论文类型：期刊论文

【摘要】：本试验采用组织块贴壁法对初生仔猪的耳组织进行原代培养,成功分离出转pGH基因猪与非转基因猪成纤维细胞,并对细胞进行形态学观察,冻存前和复苏后细胞活力检测,生长动力学分析,波形蛋白免疫组化,染色体计数以及微生物污染检测等生物学特性分析。结果表明,原代细胞经过胰蛋白酶消化和差速离心分离培养出成纤维细胞,7组细胞冻存前细胞活力均在为92%以上,冻存3个月后细胞复苏活力仍在89%以上;细胞生长总趋势呈"S"型,即经历了潜伏期、指数生长期和平台期3个阶段;细胞波形蛋白免疫组化在成纤维细胞中呈阳性反应;染色体计数结果表明,染色体数量稳定;成纤维细胞的细菌、真菌、病毒和支原体检测均为阴性。本试验成功建立了转pGH基因猪与非转基因猪成纤维细胞系。
[Abstract]:The primary culture of the ear tissue of the newborn piglets was carried out by tissue mass adherence method. The fibroblasts were isolated successfully from pigs with pGH gene and non-transgenic pigs. The morphology of the cells was observed and the viability of the cells before and after cryopreservation and resuscitation was detected. Growth kinetics analysis, vimentin immunohistochemistry, chromosome count and microbial contamination analysis. After trypsin digestion and differential centrifugation, the viability of fibroblasts in the 7 groups was above 92% before freezing, and the resuscitation activity was still over 89% after 3 months of cryopreservation, and the general trend of cell growth was "S" type. The results of chromosome count showed that the number of chromosomes was stable, the number of chromosomes was stable, the bacteria and fungi of fibroblasts, Both virus and mycoplasma were negative. In this experiment, pGH gene transgenic porcine and non-transgenic pig fibroblast cell lines were successfully established.
【作者单位】：青岛农业大学动物科技学院;
【基金】：国家转基因生物新品种培育科技重大专项(2016ZX08006-003) 山东省现代农业产业技术体系生猪创新团队建设项目(SDAIT-08-13)
【分类号】：S828

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