猪原始生殖细胞的分离_培养及鉴定.pdfVIP

动物学报 　49 (4) :529～533 , 2003 A ct a Zoolog ica S i nica 　　　　　 　 猪原始生殖细胞的分离 、培养与鉴定 ① ② ① ① ① 李世杰 　郑瑞珍 　岳奎忠 　陈　瑛 　杨增明 ( ①东北农业大学生命科学学院 , 哈尔滨 150030) ( ② 中国科学院遗传与发育生物学研究所 , 北京 100080) Isolation , culture and characterization of porcine primordial germ cell s ① ② ① ① ① L I ShiJ ie 　ZHEN G RuiZhen 　YU E KuiZhong 　CHEN Ying 　YAN G ZengMing ( ①College of L if e S ciences , N ort heast A g ricult u ral University , Harbi n 　150030 , Chi na) ( ②I nstit ute of Genetics an d Develop mental B iology , The Chi nese A cademy of S ciences , B eij i ng 　100080 , Chi na) Abstract 　Embryonic germ cells ( E G cells) are pluripotential undifferentiated stem cells isolated from cultured primordial germ cells ( P GCs) . Like ES cells , E G cells are of importance for gene targeting , t herap eutical cloning and organ trans plantation . The aim of t his study was to isolate and characterize E G cells from porcine P GCs. The genital ridges from 24 - 26 days old porcine embryos were treated in 0 02 % ED TA for 20 min and pricked wit h a needle to release P GCs. The isolated P GCs were cultured on a SNL feeder layer in an E G cell medium . The E G cell medium consisted of Dulbecco ’s modified Eagle ’s medium (DM EM) supplemented wit h 20 % Buffalo rat lever (BRL) cellconditioned medium , 15 % fe tal bovine serum , 1 mmol Lglutamine , 0 1 mol nonessential amino acids , 10 μ β ptoet hanol and antibiotics.