RRC ID 58742
著者 Naito H, Wakabayashi T, Ishida M, Gil CH, Iba T, Rahmawati FN, Shimizu S, Yoder MC, Takakura N.
タイトル Isolation of tissue-resident vascular endothelial stem cells from mouse liver.
ジャーナル Nat Protoc
Abstract Endothelial cells (ECs) are fundamental components of the blood vessels that comprise the vascular system; facilitate blood flow; and regulate permeability, angiogenesis, inflammatory responses and homeostatic tissue maintenance. Accumulating evidence suggests there is EC heterogeneity in vivo. However, isolation of fresh ECs from adult mice to investigate this further is challenging. Here, we describe an easy and reproducible protocol for isolation of different types of ECs and CD157+ vascular-resident endothelial stem cells (VESCs) by mechano-enzymatic tissue digestion followed by fluorescence-activated cell sorting. The procedure was established on liver tissue but can be used to isolate ECs from other organs with minimal modification. Preparation of single-cell suspensions can be completed in 2.5 h. We also describe assays for EC clonal and network formation, as well as transcriptomic analysis of isolated ECs. The protocol enables isolation of primary ECs and VESCs that can be used for a wide range of downstream analyses in vascular research.
巻・号 15(3)
ページ 1066-1081
公開日 2020-3-1
DOI 10.1038/s41596-019-0276-x
PII 10.1038/s41596-019-0276-x
PMID 32005982
MeSH Animals Cytological Techniques / methods* Endothelial Cells / physiology* Liver / cytology* Mice Stem Cells / physiology*
IF 10.419
引用数 0
リソース情報
ヒト・動物細胞 OP9(RCB1124)