Establishment and characterization of a fibroblast line from Simmental cattle

文献类型： 外文期刊

第一作者： Li, Lin-feng

作者： Li, Lin-feng;Guan, Wei-jun;Ma, Yue-hui;Yue, Hua;Ma, Jianzhang

作者机构：

关键词： Simmental cattle;Fibroblast line;Establishment identification

期刊名称：CRYOBIOLOGY （ 影响因子：2.487； 五年影响因子：2.641 ）

ISSN： 0011-2240

年卷期： 2009 年 59 卷 1 期

页码：

收录情况： SCI

摘要： A fibroblast line (named SCF36) from ear marginal tissue of Simmental cattle was established successfully by direct culture of explants and cell cryopreservation techniques. Biological analysis showed that the population doubling time of the thawed cells was 42.8 h. The average viability of the cells was 96.8% before freezing and 91.5% after thawing. Measurements of lactic dehydrogenase and malic dehydrogenase isoenzymes showed no cross-contamination of this cell line with other species. Karyotyping showed that the frequency of cells with chromosome number 2n = 60 was more than 90%. Tests for bacteria, fungi, viruses and mycoplasmas were negative. The efficiencies of expression of enhanced green, yellow and red fluorescent protein genes (pEGFP-N(3), pEYFP-N(1) and pDsRed1-N(1)) were between 11.3% and 28.8% after transfection; fluorescence was well distributed in the cytoplasm and nucleus except for some cryptomeric vesicles. This Simmental cattle fibroblast line not only contains the germline of this important cattle breed, which is preserved at the cellular level, but valuable material has also been provided for genomic, postgenomic and somatic cloning research. Moreover, the establishment of these methods may provide both technical and theoretical support for preserving the genetic resources of other livestock and poultry at the cellular level. (C) 2009 Elsevier Inc. All rights reserved.

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