摘要
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养体系。
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 structure was integral and the edge was obvious,Oval nucleus was located close to the central cytoplasm.Additionally,growth curve showed that GCs entered the logarithmic growth phase after 24h,and reached the highest density on the 96h,and then entered the plateau.Conclusion： cultured GCs grew well and purity 〉97%,so it is a ideal cell culture system suitable for the study of reproduction toxicity toxins.
出处
《中国兽医杂志》
CAS
北大核心
2011年第4期21-22,98,共2页
Chinese Journal of Veterinary Medicine
