兔主动脉内皮细胞的分离培养及生物学特性

Isolation and culture of rabbit aortic endothelial cells and biological characteristics

背景:为获得大量高纯度的内皮细胞,以便更好地开展对内皮细胞相关疾病的实验研究,此次研究通过改进消化酶配比比例,更新内皮细胞的分离、培养方法。目的:探索兔主动脉内皮细胞的体外培养方法,并对其进行鉴定,为更好地开展对内皮细胞相关疾病的实验研究奠定基础。方法:采用复合酶消化法,获取兔主动脉内皮组织及细胞悬液,经密度梯度离心后,获得纯度较高的血管内皮细胞,并进行体外培养。进一步对兔主动脉内皮细胞进行CD34免疫细胞化学染色鉴定,并进行细胞纯度分析。结果与结论:①体外培养出的血管内皮细胞在形态上呈典型的“铺路石”状排列;②此外免疫组化显示分离的细胞可强阳性表达CD34,证明其为内皮细胞;③对免疫组化照片进行定量分析,显示所有细胞均为CD34阳性细胞,即获得的内皮细胞纯度约大于95%;④提示采用复合酶消化法获取的兔主动脉内皮组织及细胞悬液,经密度梯度离心后,可获得纯度较高的血管内皮细胞,并通过体外培养的方法成功分离并培养了兔主动脉内皮细胞。

BACKGROUND:To obtain a large number of high-purity endothelial cells,so as to carry out better experimental research on endothelial cell-related diseases,this paper updated the separation and culture methods of endothelial cells by improving the ratio of digestive enzymes.OBJECTIVE: To explore the culture method of rabbit aortic endothelial cells in vitro and identify them, so as to lay a foundation for better experimental study ofendothelial cell-related diseases.METHODS: The endothelial tissue and cell suspension of rabbit aorta were obtained by compound enzyme digestion. After density gradient centrifugation,high-purity vascular endothelial cells were obtained and cultured in vitro. Furthermore, the rabbit aortic endothelial cells were identified by CD34immunocytochemical staining and the cell purity was analyzed.RESULTS AND CONCLUSION: (1) The vascular endothelial cells cultured in vitro were arranged in the shape of typical “paving stone”. (2) In addition,immunohistochemistry showed that the isolated cells were strongly positive for CD34-staining, to prove that they were endothelial cells. (3) The quantitativeanalysis of the immunohistochemical photos showed that all the cells were CD34-positive cells;that is, the purity of the obtained endothelial cells was morethan 95%. (4) The endothelial tissue and cell suspension of rabbit aorta were obtained by compound enzyme digestion. After density gradient centrifugation,high-purity vascular endothelial cells were obtained. Rabbit aortic endothelial cells were isolated and cultured successfully in vitro.