Abstract: Objective The experiment make a pilot study about the technique of isolating pluripotent stem cells from primordial germ cells(PGCs).Methods Fetus was derived from pregnant WuZhiShan inbreeding pigs in 26～28 d.We got fetal germ ridge,then plated on STO cells.The culture medium is composed of DMEM,2 mmol/mL L-glutamine,0.1 mmol/mL non-essential amino acid,0.1 mmol/mL β-mercaptoethanol,100 IU/mL penicillin,100 ug/mL streptomycin and 15% FCS.The embryonic germ(EG)cells was cultured and passaged.Results and Conclusions The EG cells was passaged from F_0 to F_4 or F_5.The morphology of EG cells colonies are multiplicity.By means of Alkaline phosphatase histochemical staining,Expression of stage-specific embryonic antigen-1(SSEA-1)and expression of transcription factor Oct-4 were positive.We identified that these cells possess the characteristics of embryonic stem cells.

Key words: Embryonic germ(EG)cells, Primordial germ cells(PGCs), WuZhiShan inbreeding pigs

摘要： 目的对从猪的原始生殖细胞分离获得胚胎干细胞的方法进行初步研究。方法胎儿取自怀孕26～28d的实验用五指山母猪。分离胎儿生殖嵴获得PGCs,接种于STO饲养层上,以DMEM+2mmol/L谷氨酰胺+0.1mmol/L非必需氨基酸+0.1mmol/Lβ-巯基乙醇+100IU/mL青霉素+100μg/mL链霉素+15%FCS作为培养基,对细胞进行培养传代。结果培养的细胞传至4～5代,形态多样,经AKP染色、SSEA-1免疫荧光染色、Oct-4免疫荧光标记等方法鉴定为阳性,具有胚胎干细胞的特征。

关键词: 胚胎生殖细胞, 原始生殖细胞, 五指山小型猪