Isolation of Highly Purified and Viable Retinal Endothelial Cells.
Isolation of Highly Purified and Viable Retinal Endothelial Cells.
复制标题
DOI：
10.1159/000510533
复制
发表时间：
2021
影响因子：
1.7
通讯作者：
Hirschi KK
中科院分区：
文献类型：
--
作者：
Chavkin NW;Walsh K;Hirschi KK
新生小鼠视网膜血管化模型被广泛应用于血管生物学领域，用于研究血管形成和成熟过程中血管生成和动静脉命运规范的机制。新一代测序的最新进展可以进一步阐明血管形成和重塑的机制，以及其他血管发育模型。然而，下一代测序应用需要一种分离视网膜内皮细胞的优化方法，以限制组织消化诱导的细胞损伤。在这些研究中，我们建立了一种分离新生儿视网膜内皮细胞的方法，优化了细胞活力和纯度。从消化的出生后视网膜中分离出CD31+/CD45−内皮细胞群，发现内皮细胞基因表达高度富集，并且在facs后60分钟内存活率没有变化。因此，这种分离视网膜内皮细胞的方法与下一代测序应用是兼容的。将这种分离方法与下一代测序相结合，将能够进一步描述血管发育和成熟的机制。
The neonatal mouse retinal vascularization model has been widely used in the vascular biology field to investigate mechanisms of angiogenesis and arterial-venous fate specification during blood vessel formation and maturation. Recent advances in next-generation sequencing can further elucidate mechanisms of blood vessel formation and remodelling in this, as well as other, vascular development models. However, an optimized method for isolating retinal endothelial cells that limits tissue digestion-induced cell damage is required for next-generation sequencing applications. In these studies, we established a method for isolating neonatal retinal endothelial cells that optimizes cell viability and purity. The CD31+/CD45− endothelial cell population was FACS-isolated from digested postnatal retinas, found to be highly enriched for endothelial cell gene expression, and exhibited no change in viability for 60 min post-FACS. Thus, this method for retinal endothelial cell isolation is compatible with next-generation sequencing applications. Combining this isolation method with next-generation sequencing will enable further delineation of mechanisms underlying vascular development and maturation.
登录
查看更多内容
影响因子：
23.9
作者：
Chen, Qi;Liu, Yang;Adams, Ralf H.
通讯作者：
Adams, Ralf H.
DOI：
10.1007/978-1-4939-9240-9_2
发表时间：
2019-01-01
期刊：
SINGLE CELL METHODS: SEQUENCING AND PROTEOMICS
影响因子：
--
作者：
Braga, Felipe A. Vieira;Miragaia, Ricardo J.
通讯作者：
Miragaia, Ricardo J.
DOI：
10.1007/978-1-4939-3628-1_4
发表时间：
2016-01-01
期刊：
ANGIOGENESIS PROTOCOLS, 3RD EDITION
影响因子：
--
作者：
Hewett, Peter W.
通讯作者：
Hewett, Peter W.
影响因子：
16.6
作者：
Fang JS;Coon BG;Gillis N;Chen Z;Qiu J;Chittenden TW;Burt JM;Schwartz MA;Hirschi KK
通讯作者：
Hirschi KK
影响因子：
30.8
作者：
Grosselin, Kevin;Durand, Adeline;Gerard, Annabelle
通讯作者：
Gerard, Annabelle