Establishment and biological characterization of fibroblast cell line from the Langshan chicken
Establishment and biological characterization of fibroblast cell line from the Langshan chicken
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DOI：
10.1111/j.1365-2184.2010.00666.x
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发表时间：
2010-04-01
影响因子：
8.5
通讯作者：
Ma, Y. -H.
中科院分区：
文献类型：
--
作者：
Guan, W.;He, X.;Ma, Y. -H.
目的：建立狼山鸡胚胎成纤维细胞系（LSCEF 61），从细胞水平保存狼山鸡重要的遗传资源。检测细胞形态、动态增殖和存在的任何污染，并分析染色体核型和乳酸脱氢酶和苹果酸脱氢酶同工酶水平。将pEGFP-C-1、pEGFP-N-3、pEYFP-N-1和pDsRed 1-N-1四种荧光蛋白基因转染细胞。细胞生长曲线呈典型的S形。微生物污染检测呈阴性。通过同工酶分析评估时，LSCEF 61系未显示交叉污染。染色体数目（2n）在90%以上的情况下= 78。结论：该细胞系符合美国典型培养物保藏中心的各项质量控制标准。它不仅在细胞水平上保存了重要狼山鸡的遗传资源，而且为基因组、后基因组和体细胞克隆研究等应用提供了宝贵的材料。
Objective:We needed to establish an embryonic fibroblast cell line from the Langshan chicken (LSCEF61) to preserve their important genetic resources at the cellular level.Material and methods:The cell line was established from 9-day-old embryos by direct explant culture and cryopreservation techniques. Cell morphology, dynamic proliferation and any contamination present were tested, and the karyotype and levels of isoenzymes of lactic dehydrogenase and malic dehydrogenase were analysed. Four types of fluorescent protein exogenous genes for pEGFP-C-1, pEGFP-N-3, pEYFP-N-1 and pDsRed1-N-1 were transfected into the cells.Results:Showed that the cells were healthy and were of spindle shaped structure, without change in morphology. Cell growth curves were of typical S-shape. Assays for microbial contamination were negative. The LSCEF61 line showed no cross-contamination when assessed by isoenzyme analysis. Chromosome number (2n) = 78 on more than 90% of occasions. The four types of fluorescent protein extro-genes appeared to be expressed effectively with high transfection efficiency between 15.6% and 38.6%.Conclusion:The cell line met each of the quality control standards required for the American Type Culture Collection. It had not only preserved the genetic resources of the important Langshan chicken at the cellular level, but also provided valuable material for genomic, post-genomic and somatic cell cloning research and other applications.