|本期目录/Table of Contents|

[1]张煜 李宁 白一帆 刘晓红 龚德军 徐志云.猪主动脉瓣内皮细胞的分离与鉴定[J].国际心血管病杂志,2020,01:44-47.
　ZHANG Yu,LI Ning,BAI Yifan,et al.Isolation and identification of endothelial cells from porcine aortic valve[J].International Journal of Cardiovascular Disease,2020,01:44-47.
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猪主动脉瓣内皮细胞的分离与鉴定(PDF)

《国际心血管病杂志》[ISSN:1006-6977/CN:61-1281/TN]

期数:
2020年01期
页码:
44-47
栏目:
基础研究
出版日期:
2020-01-14

文章信息/Info

Title:
Isolation and identification of endothelial cells from porcine aortic valve
作者:
张煜 李宁 白一帆 刘晓红 龚德军 徐志云
200433上海,海军军医大学长海医院心胸外科
Author(s):
ZHANG Yu;  LI Ning;  BAI Yifan;  LIU Xiaohong;  GONG Dejun;  XU Zhiyun
Department of Cardiac Surgery, Changhai Hospital, Naval Military Medical University, Shanghai200433, China
关键词:
主动脉瓣内皮细胞;  细胞培养;  细胞表型鉴定
Keywords:
Aortic valve endothelial cells;  Cell culture;  Cell phenotype identification
分类号:
-
DOI:
10.3969/j.issn.1673-6583.2020.01.011
文献标识码:
-
摘要:
目的:探讨胶原酶消化法分离猪主动脉瓣内皮细胞的可靠性及可重复性。方法:通过胶原酶消化法从新鲜猪主动脉瓣膜表面分离并培养主动脉瓣内皮细胞,通过免疫荧光染色法和流式细胞仪对获得的瓣膜内皮细胞(VEC)的表型加以鉴定。结果:通过胶原酶消化法可成功分离VEC,光镜下VEC呈铺路石样分布,为典型的内皮细胞形态。免疫荧光染色示VEC的CD31表达为阳性,波形蛋白(Vimentin)、α-平滑肌肌动蛋白(α-SMA)表达为阴性,分离获得的细胞中VEC阳性率高,间质细胞污染率较低; 流式细胞仪示分离获得的细胞Vimentin表达阴性(3.06%),CD31表达阳性(99.01%),再次验证分离获得的VEC纯度高。结论:胶原酶消化法可从猪主动脉瓣膜分离获得纯度较高的VEC,该方法简单,可重复性好。
Abstract:
Objective:To investigate the reliability and reproducibility of isolating endothelial cells from porcine aortic valve by collagenase digestion.Methods:The aortic valve endothelial cells(VECs)were isolated and cultured from fresh porcine aortic valve surface by collagenase digestion. Immunofluorescence staining and flow cytometry were used to identify the phenotype of VECs.Results:The cells isolated presented typical morphology of endothelial cells,arranging as paving stones under light microscopy. VEC could be successfully isolated by collagenase digestion. Immunofluorescence staining showed that CD31 was positive, while Vimentin and α-SMA were negative in VECs, indicating that positive rate of VECs in cells isolated was high and contamination rate by interstitial cells was low. It was verified by flow cytometry, which showed that Vimentin positive rate was 3.06% and CD31 positive rate was 99.01% in cells isolated.Conclusions:Collagenase digestion can be used to isolate aortic high-pure VECs from porcine aortic valve. This method is simple and reproducible.

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备注/Memo

备注/Memo:
基金项目:国家自然科学基金(81570351)
作者单位:200433上海,海军军医大学长海医院心胸外科
通信作者:徐志云,E-mail:zhiyunx@hotmail.com
更新日期/Last Update: 2020-01-15