Isolation and culture of rat aortic endothelial cells in vitro: A novel approach without collagenase digestion
Isolation and culture of rat aortic endothelial cells in vitro: A novel approach without collagenase digestion
复制标题
大鼠主动脉内皮细胞的体外分离和培养：一种无需胶原酶消化的新方法
DOI：
10.1002/jcb.28688
复制
发表时间：
2019-08
影响因子：
4
通讯作者：
Li Chun-Lin✳
中科院分区：
文献类型：
--
作者：
Zhu Xiao-Xiao;Miao Xin-Yu;Gong Yan-Ping;Fu Bo;Li Chun-Lin✳
为了研究心血管疾病，功能性内皮细胞的分离和培养非常重要。这项研究发现了一种新型方法来分离和培养内皮细胞。胸主动脉是从Wistar老鼠的Atta收集的
To study cardiovascular diseases, the isolation and culture of functional endothelial cells are very important. This study uncovered a novel approach to isolate and culture endothelial cells. The thoracic aorta was collected from Wistar rats with the attached tissue clearly removed. These aorta segments were seeded onto a six‐welled plate with the endothelium facing down and removed 2 days after endothelial sprouting started. The endothelial cells were harvested until 80% uneven confluence and cultured for another two passages for use in the following assays: immunofluorescence and flow cytometry assays for endothelial marker expression (CD31 and von Willebrand factor [vWF]), the Dil‐labeled acetylated low‐density lipoprotein (Dil‐Ac‐LDL) uptake assay, the tube formation assay, the Hoechst staining apoptosis assay, the β‐galactosidase staining assay for cell senescence, and the Cell Counting Kit‐8 (CCK‐8) assay for cell viability. Morphologically, the endothelial cells started to migrate away from the aorta after 50 to 72 hours of culture, showing a cobblestone‐like structure. The cultured cells expressed high levels of CD31 and vWF, 94.65% of the cells were positive for CD31, and most of the cells showed low‐density lipoprotein uptake. They were able to form tube‐like structures in vitro and were negatively stained for β‐galactosidase or Hoechst staining. Importantly, the cells at passages 3 and 10 showed similar levels of CCK‐8, β‐galactosidase, Hoechst staining, uptake of Dil‐Ac‐LDL, and capillary tube formation. This novel technique is useful to isolate and culture rat aortic endothelial cells for future studies of endothelial functions and biology. In addition, primary vascular endothelial cells at passages 3 to 10 are suitable for experiments.
登录
查看更多内容
影响因子：
4.4
作者：
Yamagishi SI
通讯作者：
Yamagishi SI
影响因子：
5.2
作者：
Steve George
通讯作者：
Steve George
DOI：
10.1016/s1051-0443(98)70321-5
发表时间：
1998-07
影响因子：
2.9
作者：
J. Mcgraw
通讯作者：
J. Mcgraw
影响因子：
--
作者：
Montserrat Fibla;Anna Hernández-Aguilera;J. Camps;J. Menéndez;J. Joven
通讯作者：
Montserrat Fibla;Anna Hernández-Aguilera;J. Camps;J. Menéndez;J. Joven
影响因子：
5.9
作者：
Caplan, Louis R.;Searls, D. Eric;Hon, Fong Kwong Sonny
通讯作者：
Hon, Fong Kwong Sonny