| 猪卵巢颗粒细胞分离培养及鉴定 |
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| 引用本文： | 朱丽,郭成志,邓思君,鲁银,魏强,袁慧. 猪卵巢颗粒细胞分离培养及鉴定[J]. 中国兽医杂志, 2011, 47(4) |
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| 作者姓名： | 朱丽 郭成志 邓思君 鲁银 魏强 袁慧 |
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| 作者单位： | 湖南农业大学动物医学院,湖南长沙,410128 |
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| 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.
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| 关 键 词： | 猪 卵巢颗粒细胞 细胞培养 免疫细胞化学 |
Isolation, culture and identification of porcine ovary granulose cells |
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| ZHU Li,GUO Cheng-zhi,DENG Si-jun,LU Yin,WEI Qiang,YUAN Hui. Isolation, culture and identification of porcine ovary granulose cells[J]. Chinese Journal of Veterinary Medicine, 2011, 47(4) |
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| Authors: | ZHU Li GUO Cheng-zhi DENG Si-jun LU Yin WEI Qiang YUAN Hui |
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| Affiliation: | ZHU Li,GUO Cheng-zhi,DENG Si-jun,LU Yin,YUAN Hui(College of Veterinary Medicine,Hunan Agricultural University,Changsha 410128,China) |
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| Abstract: | 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 ... |
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| Keywords: | swine granulosa cells cell culture immocytochemistry |
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