期刊论文

Isolation,culture and identification of porcine ovary granulosa cells

中文

中国兽医杂志

0529-6005

2011

47

4

21-22,中插2

10.3969/j.issn.0529-6005.2011.04.008

本校为第一机构

为构建猪卵巢颗粒细胞的体外培养体系,研究毒素的生殖毒性奠定基础,采用机械分离法从1～5 mm的卵泡中提取猪卵巢颗粒细胞,苔盼蓝染色计算细胞存活率,Giemsa染色和结晶紫染色观察细胞形态,免疫细胞化学染色方法检测促卵泡刺激素受体（FSHR）表达来鉴定颗粒细胞,MTT方法测定细胞生长曲线。结果显示分离提取的猪卵巢颗粒细胞存活率为65%左右,细胞纯度>97%,细胞结构完整,边缘分明,胞核呈卵圆形位于靠近胞质的中央。另外细胞生长曲线表明,细胞从24 h开始进入对数生长期,并于96 h达到最高密度,之后进入平台期。分离培养的颗粒细胞生长状态良好,且细胞纯度>97%,是理想的适合进行毒素生殖毒理学的细胞培...

To establish a cultured system in vitro of porcine ovary granulosa cells（GCs） for the study of reproductive toxicity toxins,GCs was extracted from follicles（1-5mm） by mechanical separation.Morphology of GCs was observed by a microscope using Giemsa and crytal violet staining.Cell viability was determined by 0.4% trypan blue.Exclusion immunocytochemistry was used to measure the FSHR expression to characterize GCs.In addition,Growth curve of GCs was detected by MTT assay.The results showed that GCs viability was about 65%,purity〉 97%.GCs str...