| 原代人脐静脉内皮细胞分离培养方法的探讨及其鉴定 |
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| 引用本文： | 严凤英,于萍,关林波,彭冰,周淑. 原代人脐静脉内皮细胞分离培养方法的探讨及其鉴定[J]. 华西医学, 2010, 0(4): 727-729 |
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| 作者姓名： | 严凤英 于萍 关林波 彭冰 周淑 |
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| 作者单位： | [1]成都市计划生育指导所,成都610031 [2]四川大学华西第二医院细胞与基因治疗实验室 ,成都610031 [3]四川大学华西第二医院妇产科,成都610031 |
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| 基金项目： | 国家自然科学基金资助项目（30600677）; 教育部留学回国人员科研启动基金[2009]8-8-5 |
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| 摘 要： | 目的通过比较两种原代人脐静脉内皮细胞的分离培养方法并对细胞特异性抗原进行鉴定,探索提高原代内皮细胞体外培养存活率及纯化率的方法。方法采用一次性无菌注射器向人脐静脉灌注消化液,消化液的浓度和消化时间分别0.25%（质量体积比）胰蛋白酶,10min和0.1%（质量体积比）胶原酶Ⅱ,15min。通过在倒置显微镜下观察细胞的形态特点和用免疫荧光染色的方法对细胞进行鉴定,比较两种消化方法的优劣。结果 0.1%胶原酶Ⅱ,15min的消化方法较0.25%胰蛋白酶,10min对原代人脐静脉内皮细胞有更好的分离效果,活细胞数量多且细胞纯度较高。免疫荧光染色结果表明细胞内有Ⅷ因子相关抗原表达。结论胶原酶Ⅱ可以有效分离脐静脉内皮细胞,最佳消化条件是0.1%胶原酶Ⅱ,37℃,15min。
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| 关 键 词： | 人脐静脉内皮细胞 分离 培养 |
Isolation and Identification of Primary Human Umbilical Vein Endothelial Cells |
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| YAN Feng-ying,YU Ping,GUAN Lin-bo,PENG Bing,ZHOU Shu. Isolation and Identification of Primary Human Umbilical Vein Endothelial Cells[J]. West China Medical Journal, 2010, 0(4): 727-729 |
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| Authors: | YAN Feng-ying YU Ping GUAN Lin-bo PENG Bing ZHOU Shu |
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| Affiliation: | 1.Chengdu Institute of Family Planning,Chengdu,Sichuan,610031,P.R.China;2.Laboratory of Cell and Gene Therapy;3.Department of Pediatrics,West China Second Hospital of Sichuan University,Chengdu,Sichuan 610041,P.R.China ) |
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| Abstract: | Objective To explore the optimal method for primary culture of human umbilical vein endothelial cells（HUVECs）.Methods HUVECs were prepared from human umbilical cords by 0.1% collagenase Ⅱ digestion for 15 minutes and 0.25 trypsinase digestion for 10 minutes,respectively.HUVECs were observed under inverted microscope and identified by immunofluorescence.The two methods of digestion were compared.Results More HUVECs were harvested through the method of 0.1% collagenase Ⅱ for 15 minutes,which expressed Ⅷ related antigen.Conclusion The method of 0.1% collagenase Ⅱ digestion for 15 minutes is a better choice to isolate HUVECs. |
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| Keywords: | Human umbilical vein endothelial cells Seperation Culture |
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