蒙古绵羊原始生殖细胞的分离、培养及鉴定

Isolation, culture, and characterization of primordial germ cells in Mongolian sheep.

作者信息

Liu Chun Xia, Wang Wen Long, Zhao Rui Yuan, Wang Hai Tao, Liu Yi Yi, Wang Shen Yuan, Zhou Huan Min

机构信息

College of Life Science, Inner Mongolia Agricultural University, 306 Zhao wu da street, Hohhot, Inner Mongolia, 010018, China.

出版信息

In Vitro Cell Dev Biol Anim. 2014 Mar;50(3):207-13. doi: 10.1007/s11626-013-9695-0. Epub 2013 Oct 19.

DOI:10.1007/s11626-013-9695-0

PMID:24142152

Abstract

This study was performed to culture and preliminarily identify the primordial germ cells (PGCs) isolated from the genital ridge of the Mongolian sheep fetus. The growth characteristics of the sheep PGCs were detected in different culture systems such as culture media, resources, and state and passages of feeder cells. The obtained embryonic germ (EG) cells were identified by morphology, enzymology, and immunofluorescence. The results showed that the sheep EG cell colonies were ridgy, typically nest like, and compact, and had regular edges. Alkaline phosphatase staining reaction was weakly positive. EG cells expressed Kit, Rex-1, Nanog, and Oct-4. Immunofluorescence detection was weakly positive for Oct3/4, whereas positive for SSEA-1, SSEA-3, SSEA-4, TRA-1-61, and TRA-1-80.

摘要

本研究旨在培养并初步鉴定从蒙古绵羊胎儿生殖嵴分离出的原始生殖细胞（PGCs）。在不同培养体系中检测绵羊PGCs的生长特性，如培养基、资源以及饲养层细胞的状态和传代情况。通过形态学、酶学和免疫荧光法对获得的胚胎生殖（EG）细胞进行鉴定。结果显示，绵羊EG细胞集落呈嵴状，典型的巢状且紧密，边缘规则。碱性磷酸酶染色反应呈弱阳性。EG细胞表达Kit、Rex-1、Nanog和Oct-4。免疫荧光检测Oct3/4呈弱阳性，而SSEA-1、SSEA-3、SSEA-4、TRA-1-61和TRA-1-80呈阳性。