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Department of Molecular Biology and Biochemistry, University of California, Irvine (UCI)
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Davis, J., Crampton, S. P., Hughes, C. C. Isolation of Human Umbilical Vein Endothelial Cells (HUVEC). J. Vis. Exp. (3), e183, doi:10.3791/183 (2007).
血管生成是一个复杂的多步骤的过程,在血管生成刺激,从现有的血管创建新船。这些步骤包括:降解基底膜,增殖和迁移(发芽)内皮细胞到细胞外基质,对齐欧共体(EC)的成线,形成管腔吻合,并形成一个新的基底膜。在体外实验中有许多已开发研究这个过程,但大多只模仿血管生成的某些阶段,形态的船只往往并不像在体内的船只。在这里,我们证明,利用人脐静脉EC和成纤维细胞在体外血管生成试验进行了优化。这种模式概括所有血管生成的关键早期阶段,而重要的血管显示专利包围极化欧共体间流明。船只,可以很方便地观察相衬和时间推移显微镜,并在下游应用纯粹的形式收回。
程序
注:(未激活)的内皮细胞有鹅卵石的外观。有时激活内皮细胞(长,尖尖的)后隔离,后一对夫妇通过他们通常会返回到一个unactive状态。
净化
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| Name | Company | Catalog Number | Comments |
| 0.1% Collagenase | 10ml per cord, warmed to 37°C | ||
| Tissue culture flasks | T75, one per cord. | ||
| Hanks medium | |||
| scissors | sterile | ||
| beaker | sterile | ||
| 50 ml tubes |
may you tell me wath kind of collagenase do you use? thank you!
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ReplyPosted by: loryMay 16, 2008, 5:47 AM