Lentiviral vector transfection of chicken embryo primordial germ cells in vitro

Ding Hong-mei, Xu Shi-yong, Shao Gen-bao, Sun Yan, Wang Meng, Liu Honglin
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引用次数: 1

Abstract

Lentiviruses as gene transfer vectors have been used successfully to transfect mammal embryonic stem cells and germline stem cells, but this has not been attempted in avian primordial germ cells (PGCs). PGCs were isolated from the gonads of Isa-brown chicken embryos at stage 28 and co-cultured with gonadal stroma cells. A lentiviral vector pLenti-CMV-EGFP was constructed and the virus harvested by cotransfecting 293FT cells with the vector and packaging plasmids. Concentrated lentiviruses were used to transfect chicken PGCs, the transfection efficiency was up to 24.19%.
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慢病毒载体体外转染鸡胚原始生殖细胞的研究
慢病毒作为基因转移载体已成功转染哺乳动物胚胎干细胞和种系干细胞,但尚未尝试转染禽类原始生殖细胞(PGCs)。从28期伊沙褐鸡胚性腺中分离出PGCs,并与性腺基质细胞共培养。构建慢病毒载体plentiv - cmv - egfp,用载体和包装质粒共转染293FT细胞,收获病毒。采用浓缩慢病毒转染鸡PGCs,转染效率可达24.19%。
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