四川农业大学学报

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兔血管内皮细胞的体外培养及生物学鉴定

王文秀,沈志强   

  1. 山东省滨州畜牧兽医研究院
  • 收稿日期:2011-09-15 修回日期:2011-11-27 出版日期:2011-12-25 发布日期:2011-12-26
  • 通讯作者: 沈志强

Culture and Biological Characterastics of Rabbit Vascular Endothelial Cells

  1. 1. Shandong Binzhou Animal Science & Veterinary Medicine Institute
    2.
  • Received:2011-09-15 Revised:2011-11-27 Online:2011-12-25 Published:2011-12-26

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,约一周长成单层,纯度较高;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 primary culture of rabbit vascular endothelial cells (RVECs) in vitro was to study the pathogenicity of rabbit haemorrheagic disease virus on RVECs. To culture RVECs by trypsin-digest method and tissue pieces-stick method and identify the cells by factor Ⅷ related antigen immunohistochemical staining and CD34 indirect immunofluorescence assay. The result showed that cells isolated by trypsin-digest method adhered about 20 h, 1 week later, constituted a confluent monolayers. Cells were passaged, they constituted a confluent monolayer cells and most just like polygon and spindle-shaped. Immunohistochemical staining and indirect immunofluorescence revealed that the expression of factor Ⅷ and CD34 was positive. By tissue pieces-stick method, the cells grew form tissues about 2 weeks and constituted a confluent monolayer cells by third week. These cells had higher activity but they were contaminated with smooth muscle cells and fibroblast cells.

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