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Study on characteristics of in vitro culture and intracellular transduction of exogenous proteins in fibroblast cell line of Liaoning cashmere goat

文献类型： 外文期刊

作者： Hu, P. F. 1 ; Guan, W. J. 1 ; Li, X. C. 1 ; Zhang, W. X. 1 ; Li, C. L. 1 ; Ma, Y. H. 1 ;

作者机构： 1.Chinese Acad Agr Sci, Inst Anim Sci, Beijing 100193, Peoples R China

2.Liaoning Acad Agr Sci, Inst Wind Sandy Land Improvement & Utilizat, Fuxing 123000, Peoples R China

关键词： Liaoning cashmere goat; Fibroblast cell line; Exogenous protein; TAT peptide; Transduction

期刊名称：MOLECULAR BIOLOGY REPORTS （ 影响因子：2.316； 五年影响因子：2.357 ）

ISSN： 0301-4851

年卷期： 2013 年 40 卷 1 期

页码：

收录情况： SCI

摘要： Establishment of fibroblast cell lines of endangered goat breeds and research on the gene or protein functions based on the cells made a significant contribution to the conservation and utilization of genetic resources. In this study, a fibroblast cell line of Liaoning cashmere goat, frozen in 174 cryovials with 5 x 10(6) cells each, was successfully established from 60 goats ear marginal tissues using explant culture and cryopreservation techniques. Biological analysis of in vitro cultured cell line showed that, the cells were morphologically consistent with fibroblasts; the average viability of the cells was 94.9 % before freezing and 90.1 % after thawing; the growth process of cells was consisted of a lag phase, a logarithmic phase and a plateau phase; cell population doubling time was 65.5 h; more than 90 % of cells were diploid prior to the 6th generation; Neither microbial contamination nor cross-contamination was detected. To determine cell permeability, intracellular path and stability of exogenous proteins during the transduction, a TAT protein transduction domain was fused to the C-terminus of enhanced green fluorescent protein, the established fibroblast cell line was treated with the purified exogenous proteins at various concentrations by adding them to the cell culture media for 1-24 h and assayed cell morphology and protein presence, it was found that the purified exogenous proteins readily entered cells at a concentration of 0.1 mg/ml within 1.5 h and some of them could translocate into nucleus, moreover, the exogenous proteins appeared to be stable inside cells for up to 24 h.

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