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首页> 外文期刊>Animal Science Journal >Establishment and characterization of a fibroblast cell line derived from Mongolian sheep
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Establishment and characterization of a fibroblast cell line derived from Mongolian sheep

机译:蒙古绵羊成纤维细胞系的建立和鉴定

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摘要

The Mongolian sheep ear marginal tissue fibroblast cell line (MSF32) from 32 samples was successfully established by using primary explants technique and cell cryoconservation technology. MSF32 cells were adherent, with a population doubling time of 28.2?h. Chromosome analysis showed that >90.2% of cells were diploid (2n?=?54) prior to cell passage 4. Isoenzyme analyses of lactate dehydrogenase and malate dehydrogenase showed that the MSF22 cells had no cross-contamination with other species. Tests for cell line contamination with bacteria, fungi, viruses and mycoplasmas were also negative. Plasmids encoding the fluorescent proteins pEGFP-N3, pEGFP-C1, pECFP-N1, pECFP-mito, pDsRed1-N1 and pEYFP-N1 were transfected into cells to study exogenous gene expression in the cells. The plasmid transfection efficiency was between 12.3% and 63.3%. Every index of the MSF32 cell line meets all the standard quality controls of American Type Culture Collection (ATCC). Not only has the genetic resources of the Mongolian sheep been preserved at the cell level, but also valuable materials had been provided for genome, postgenome and somacloning research.
机译:应用原发外植体技术和细胞冷冻保存技术成功地从32个样本中建立了蒙古绵羊耳缘组织成纤维细胞系(MSF32)。 MSF32细胞是贴壁的,群体倍增时间为28.2?h。染色体分析显示,> 90.2%的细胞在细胞传代前是二倍体(2n?=?54)。乳酸脱氢酶和苹果酸脱氢酶的同工酶分析表明,MSF22细胞与其他物种没有交叉污染。细胞株被细菌,真菌,病毒和支原体污染的测试也为阴性。将编码荧光蛋白pEGFP-N3,pEGFP-C1,pECFP-N1,pECFP-mito,pDsRed1-N1和pEYFP-N1的质粒转染到细胞中,以研究细胞中的外源基因表达。质粒转染效率在12.3%和63.3%之间。 MSF32细胞系的每个指标均符合美国典型培养物保藏中心(ATCC)的所有标准质量控制。不仅在细胞水平上保存了蒙古绵羊的遗传资源,而且还为基因组,后基因组和体细胞克隆研究提供了有价值的材料。

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