慢病毒载体体外转染鸡胚原始生殖细胞研究

提取第28 期鸡胚性腺中的原始生殖细胞(PGCs)，将其与性腺基质细胞共培养，并对PGCs 进行PAS (过碘酸希夫试剂) 和AKP(碱性磷酸酶) 染色鉴定。构建pLenti-CMV-eGFP 慢病毒表达载体，与辅助质粒共转染293FT 细胞生产病毒颗粒。将慢病毒浓缩后转染鸡胚PGCs，转染率高达24.19 %。实验还比较了慢病毒不同剂量对转染效率的影响，随着慢病毒剂量的增加，转染效率显著提高。

Lentiviral vectors transfection of chicken primordial germ cells

Primordial germ cells (PGCs) were isolated from the gonads of chicken embryos at stage 28, PGCs were co-cultured with gonadal stroma cells. Identification of PGCs was carried out by periodic acid-Schiff (PAS) and alkaline phosphatase (AKP) staining. We constructed a lentiviral vector pLenti-CMV-eGFP and harvested the virus by cotransfecting 293FT cells with the vector and packaging plasmids. Lentiviruses after concentration were used to transfect chicken PGCs, the transfection efficiency of PGCs was up to 24.19%. The impacts of different volume of lentiviruses on transfection efficiencies were also compared, the results indicated that as the increase of lentiviruses, the transfection efficiency was significantly improved.