四川农业大学学报 ›› 2011, Vol. 29 ›› Issue (04): 544-548.doi: 10.3969/j.issn.1000-2650.2011.04.020

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兔血管内皮细胞的体外培养及生物学特性

王文秀1,2, 唐娜1,2, 谢金文1,2, 莫玲1, 庄金秋1, 张松林1, 沈志强1,2   

  1. 1. 山东省滨州畜牧兽医研究院, 山东 滨州 256600;
    2. 山东绿都生物科技有限公司, 山东 滨州 256600
  • 收稿日期:2011-09-15 出版日期:2011-12-31 发布日期:2017-04-12
  • 通讯作者: 沈志强,博士,研究员,主要从事预防兽医学与兽医生物制品研究,E-mail:zshenzq@163.com。 E-mail:zshenzq@163.com
  • 作者简介:王文秀,博士,助理研究员,主要从事分子病原学与免疫学研究,E-mail:wwx1997@126.com。
  • 基金资助:
    山东省自主创新成果转化重大专项计划(2008ZHZX1A1103)

Culture and Identification of Vascular Endothelial Cells in Rabbit

WANG Wen-xiu1,2, TANG Na1,2, XIE Jin-wen1,2, MO Ling1, ZHUANG Jin-qiu1, ZHANG Song-lin1, SHEN Zhi-qiang1,2   

  1. 1. Shandong Binzhou Animal Science and Veterinary Medicine Institute, Binzhou 256600, Shandong, China;
    2. Shandong Lvdu Bio-Sciences and Technology Co. Ltd., Binzhou 256600, Shandong, China
  • Received:2011-09-15 Online:2011-12-31 Published:2017-04-12

摘要: 为研究兔瘟病毒对兔血管内皮细胞的致病性,建立兔血管内皮细胞的体外培养方法。采用消化法和组织块贴壁法分离获得兔子血管内皮细胞,对其进行体外培养,并对细胞进行第Ⅷ因子相关抗原的免疫组织化学鉴定和CD34的间接免疫荧光检测。结果表明,消化法分离的细胞20h左右贴壁,约1周长成单层,纯度较高;组织块贴壁法分离的细胞两周才开始从组织块爬出,约3周长满单层,有杂细胞污染,但细胞活力相对较高。消化法获得的细胞传代后多呈单层贴壁生长,形状以多角形和梭状居多,传代后呈典型的"铺路石"状,第Ⅷ因子相关抗原和CD34检测均为阳性。

关键词: 细胞培养, 兔血管内皮细胞, 生物学特性

Abstract: The aim of the study was to develop a method for the isolation, identification and primary culture of rabbit vascular endothelial cells(RVECs) as a modle for studying the pathologenicity of rabbit haemorrheagic disease virus on RVECs.RVECs were dissociated by trypsin digestion and tissue pieces culture method and cultured in vitro.The REVCs were identified by factor Ⅷ related antigen immunohistochemical staining and CD34 indirect immunofluorescence assay.The result showed that the cells which gained by digesting method adhered about 20 h, and filled the basis of petri dish after 1w with high activity.The cells which isolated by tissue pieces culture method grew from original about 2 weeks and filled the basis of petri dish 3 weeks later, and these cells were contaminated with smooth muscle cells and fibroblast cells, while they had higher vitality.After cells were passaged and cultured, they exhibited as polygon and spindle-shaped cells and displayed cobblestone-likemorphology after fusion to monolayer.Immunohistochemical staining and indirect immunofluorescence revealed that the expression of factor Ⅷ and CD34 was positive.

Key words: cell culture, rabbit vascular endothelial cells, biological characteristics

中图分类号: 

  • R322.12