联系我们 加入收藏
 
年期检索 高级检索
33
2025年4月5日 星期六
  研究论文 本期目录 | 过刊浏览 | 高级检索 |
鸡胚原始生殖细胞的分离、培养及鉴定
唐新燕;曾卫东;米玉玲;刘红云;张才乔
浙江大学动物科学学院, 杭州310029
Isolation, Culture and Characterization of Chicken Primordial Germ Cells
TANG Xin-yan;ZENG Wei-dong;MI Yu-ling;LIU Hong-yun;ZHANG Cai-qiao
College of Animal Sciences, Zhejiang University, Hangzhou 310029, China
全文: PDF (305 KB)   HTML (1 KB) 
输出: BibTeX | EndNote (RIS)      
摘要 采用Ficoll密度梯度离心法分离第19期(孵化68~72 h)艾维茵鸡(Gallus domesticus)生殖嵴原始生殖细胞(PGC),通过体细胞-PGC共培养进行了原代和传代培养,对PGC进行了碱性磷酸酶(AKP)和过碘酸雪夫氏反应(PAS)鉴定,并用细胞增殖核抗原(PCNA)免疫细胞化学检测了传代于鸡胚饲养层上PGC的增殖活性。培养的PGC经AKP和PAS鉴定均为阳性,AKP阳性为红棕色,PAS阳性为深红色,PCNA染色显示PGC集落为红棕色,表明PGC处于增殖状态。同时比较了不同体外培养条件(5%~20%胎牛血清(FCS), ITS培养液(M199+ 10 μg/mL 胰岛素+5 μg/mL转铁蛋白+3×10-8 mol/L亚硒酸钠), 条件培养液, 15%FCS+ITS, 15%FCS+40% 条件培养液)下PGC增殖情况。发现未经密度梯度离心的PGC生长较离心的PGC好,且5%FCS在原代培养中起较好的促增殖作用;而传代培养时在不同培养条件下,5%FCS组或ITS组形成的PGC集落的直径较大,而仅用单纯的条件培养液效果并不明显。结果表明除鸡胚饲养层可作为PGC增殖的支持体系外,在体细胞-PGC共培养方式下5%FCS或单独的ITS也可作为PGC体外增殖的一种培养体系。
服务
把本文推荐给朋友
加入我的书架
加入引用管理器
E-mail Alert
RSS
作者相关文章
唐新燕
曾卫东
米玉玲
刘红云
张才乔
关键词 原始生殖细胞鸡胚细胞增殖体外培养    
AbstractPrimordial germ cells (PGCs) were isolated from the genital ridges of Avian chicken (Gallus domesticus) embryos at 19th stage and purified by Ficoll density-gradient centrifugation. PGCs were co-cultured with somatic cells in primary culture and subculture. Identification of PGCs was carried out by histochemical methods including alkaline phosphatase (AKP) and periodic acid-Schiff (PAS) staining, and proliferating PGCs in subculture was demonstrated by immunocytochemistry of proliferating cell nuclear antigen (PCNA). Meanwhile, proliferating activity of PGCs was compared under different culture conditions among 5% ~20% fetal calf serum (FCS), ITS (M199+10 μg/mL insulin+5 μg/mL transferrin + 3×10-8 mol/L selenite, conditioned medium (CM), 15%FCS+ITS, 15%FCS+40% CM. The results showed that the cultured PGCs were positive for AKP and PAS staining and displayed intensive proliferating activity by PCNA. PGCs grew better without centrifugation than those with centrifugation. PGCs formed larger colonies in media with 5%FCS or ITS than those in other media. The above results indicate that 5%FCS or ITS-supplemented media can be a culture system for PGC proliferation in the somatic cell- PGC coculture, besides the embryonic fibroblast feed layer.
Key wordsprimordial germ cells    chicken embryo    cell proliferation    in vitro culture
收稿日期: 2005-04-04     
ZTFLH:  S185  
通讯作者: 张才乔   
引用本文:   
唐新燕;曾卫东;米玉玲;刘红云;张才乔. 鸡胚原始生殖细胞的分离、培养及鉴定[J]. , 2006, 14(2): 174-177.
TANG Xin-yan;ZENG Wei-dong;MI Yu-ling;LIU Hong-yun;ZHANG Cai-qiao. Isolation, Culture and Characterization of Chicken Primordial Germ Cells. , 2006, 14(2): 174-177.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/      或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2006/V14/I2/174
 
版权所有 © 2014 《农业生物技术学报》编辑部   京ICP备11035905号-3
地址:北京市海淀区圆明园西路2号中国农业大学生命科学楼1053室 邮编:100193
电话:010-62733684 传真:010-62731615 E-mail: nsjxb@cau.edu.cn