Laboratory of Angiogenesis and Vascular Metabolism, Center for Cancer Biology, VIB, Department of Oncology and Leuven Cancer Institute (LKI), KU Leuven, Leuven, Belgium.
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-Sen University, Guangzhou 510060, Guangdong, P.R. China.
STAR Protoc. 2021 Jul 28;2(3):100523. doi: 10.1016/j.xpro.2021.100523. eCollection 2021 Sep 17.
Endothelial cells (ECs) exhibit phenotypic and functional tissue specificities, critical for studies in the vascular field and beyond. Thus, tissue-specific methods for isolation of highly purified ECs are necessary. Kidney, spleen, and testis ECs are relevant players in health and diseases such as chronic kidney disease, acute kidney injury, myelofibrosis, and cancer. Here, we provide tailored protocols for rapid and reproducible EC purification established for scRNA sequencing from these adult murine tissues using the combination of magnetic- and fluorescence-activated cell sorting. For complete details on the use and execution of these protocols, please refer to Kalucka et al. (2020) and Dumas et al. (2020).
内皮细胞(ECs)表现出表型和功能的组织特异性,这对于血管领域及其他领域的研究至关重要。因此,需要使用组织特异性方法来分离高度纯化的 ECs。肾脏、脾脏和睾丸 ECs 是健康和疾病(如慢性肾脏病、急性肾损伤、骨髓纤维化和癌症)中的重要参与者。在这里,我们提供了针对这些成年鼠组织的 scRNA 测序的快速且可重复的 EC 纯化方案,该方案结合了磁性和荧光激活细胞分选。有关这些方案的使用和执行的完整详细信息,请参阅 Kalucka 等人(2020 年)和 Dumas 等人(2020 年)。