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Publication : A robust and efficient microvascular isolation method for multimodal characterization of the mouse brain vasculature.

First Author  Bjørnholm KD Year  2023
Journal  Cell Rep Methods Volume  3
Issue  3 Pages  100431
PubMed ID  37056377 Mgi Jnum  J:348916
Mgi Id  MGI:7643669 Doi  10.1016/j.crmeth.2023.100431
Citation  Bjornholm KD, et al. (2023) A robust and efficient microvascular isolation method for multimodal characterization of the mouse brain vasculature. Cell Rep Methods 3(3):100431
abstractText  Studying disease-related changes in the brain vasculature is warranted due to its crucial role in supplying oxygen and nutrients and removing waste and due to the anticipated vascular dysfunction in brain diseases. To this end, we have developed a protocol for fast and simple isolation of brain vascular fragments without the use of transgenic reporters. We used it to isolate and analyze 22,515 cells by single-cell RNA sequencing. The cells distributed into 23 distinct clusters corresponding to all known vascular and perivascular cell types in the brain. Western blot analysis also suggested that the protocol is suitable for proteomic analysis. We further adapted it for the establishment of primary cell cultures. The protocol generated highly reproducible results. In conclusion, we have developed a simple and robust brain vascular isolation protocol suitable for different experimental modalities, such as single-cell analyses, western blotting, and primary cell culture.
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