Establishment and characterization of a fibroblast cell line derived from Gansu alpine fine-wool sheep

文献类型： 外文期刊

第一作者： Pei, Pei

作者： Pei, Pei;He, Xiaohong;Zhao, Yuhua;Zhu, Zhiqiang;Kan, Junchang;Guan, WeiJun;Ma, Yuehui

作者机构：

关键词： cell culture;chromosome and karyotype analysis;expression of fluorescent protein gene

期刊名称：BIOCELL （ 影响因子：1.254； 五年影响因子：0.924 ）

ISSN： 0327-9545

年卷期： 2015 年 39 卷 1 期

页码：

收录情况： SCI

摘要： Afibroblast cell line was successfully established from ear marginal tissue of Gansu alpine fine-wool sheep by using a primary explant technique and cell cryogenic preservation technology. Cryopreserved cells were checked for quality and biological characteristics and showed typical fibroblast morphology when cultured again in vitro. The growth curve was typically S-shaped as the cell population passed through a lag phase, a logarithmic phase and a plateau phase. Biological analysis showed that the population doubling time (PDT) for reviving cells was approximately 24 h. The cells were also free from bacterial, fungal, viral and mycoplasma contamination. Karyotyping analysis indicated that the chromosome number of a normal cell was of 2n = 54 and 95.4% of the entire population was diploid. The transfection efficiencies of three fluorescent proteins (pEGFP-N3, pDsRed-N1, pEYFP-N1) optimal at 48h were from 18.5% to 30.1%. The corresponding fluorescence was distributed throughout the cytoplasm and nucleus 24 h after transfection. This newly established cell line meets the quality control standards established by the American Type Culture Collection. It not only preserves the genetic resources of the Gansu alpine fine-wool sheep at the cellular level but also provides valuable materials for genomic, postgenomic, and somatic cloning research in these animals. 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.

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