| 摘要: |
| 【目的】以滩羊胎儿皮肤组织为材料，获得符合体细胞克隆转基因基本要求的供体细胞系。【方法】取滩羊胎儿皮肤组织，采用组织块法分离培养获得滩羊胎儿成纤维细胞，观测其形态和生长情况，鉴定其性别，并对其染色体核型和红色荧光报告基因质粒(pmCherry-N1)转染等生物学特性进行研究。【结果】采用组织块法成功获得了滩羊胎儿成纤维细胞，其细胞群体倍增时间（PDT）约为48 h；5代、15代、25代滩羊胎儿成纤维细胞核型分析表明，各代细胞染色体均为2n=54，未见异常；滩羊胎儿成纤维细胞的转染效率较高，约为50%，对最终获得的稳定表达红色荧光蛋白的胎儿成纤维细胞进行核型分析，结果证明遗传物质没有发生变化。【结论】建立了滩羊胎儿成纤维细胞系，为滩羊分子育种奠定了基础。 |
| 关键词: 滩羊 成纤维细胞 生物学特性 体外培养 |
| DOI： |
| 分类号: |
| 基金项目:宁夏自然科学基金项目 |
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| Establishment of fetal fibroblast cell line and its biological characteristics in Tan sheep |
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WANG Juan,SUN Yi,CHENG Long,et al
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| Abstract: |
| 【Objective】This study aimed to obtain donor cells for somatic cloning and gene transformation from fetal skin tissue of Ningxia Tan sheep.【Method】Fetal skin tissues of Ningxia Tan sheep were cultivated by explant culture technique to obtain fetal fibroblast cells.The red fluorescent protein report gene (pmCherry-N1) was transferred into fetal fibroblast cells with liposome.The morphology and dynamic growth of obtained cells were analyzed,the sex was determined,and the karyotype was studied.【Result】Fibroblast cell of Tan sheep fetal was acquired by explant culture technique with the population doubling time (PDT) of approximately 48 h.Karyotype analysis of 5,15,and 25 generations showed that the chromosome diploid was 2n=54,and no anomalies were observed.Transfer efficiency of the plasmid was 50%,and karyotype analysis of fetal fibroblast cells,which generated stably expressed red fluorescent protein report gene,showed that the hereditary character was stable.【Conclusion】Fetal fibroblast cell lines in Ningxia Tan sheep were successfully established,which would improve the breeding of Tan sheep. |
| Key words: Tan sheep fibroblast cell biological characteristics in vitro culture |
