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Identification Of A Circulating Pericyte That Homes To Ischemia
Melanie Rodrigues, PhD, Robert C. Rennert, MD, Michael Januszyk, MD, Alexander Y. Li, MS, Nigel Kooreman, MD, Christopher R. Davis, MD, Charles K. Chan, PhD, Nathaniel B. Fernhoff, PhD, Tina Ngyuen, High School, Michael Sorkin, MD, Joseph C. Wu, MD, Irving Weissman, MD, Geoffrey C. Gurtner, MD.
Stanford, Stanford, CA, USA.

PURPOSE: Pericytes are non-hematopoietic cells found adjacent to endothelial cells in blood vessels. They assist in neovascularization by scaffolding and stabilizing endothelial cells. However, the origin, identity and fate of pericytes remain unclear. Here, we test for a circulating pericyte following ischemic injury.
METHODS: Murine parabiosis is established between a fluorescent reporter mouse (donor) and a non-fluorescent syngenic mouse (recipient). Hind-limb ischemia or ischemic flaps are created on the recipient mouse following establishment of cross-circulation. The ischemic sites on the recipient mouse are isolated 7, 14, 28 or 180 days following injury. All circulating non-immune cells are isolated by fluorescent assisted cell sorting and subjected to single-cell transcriptional analysis for genes specific to stemness, neovascularization, and growth factor receptors, to determine the fate and profile of cells. Bone marrow of the reporter mouse is transplanted into non-fluorescent syngenic recipients, and ischemic flaps are made on the recipients to determine the source of the circulating cells.
RESULTS: Single-cell analysis of non-immune cells recruited to ischemia identifies a unique bone marrow derived pericyte that attaches to endothelial cells at the site of injury, deposits extra-cellular matrix, and proliferates following engraftment. We find no circulating cell that differentiates into an endothelial cell at the site of ischemia.
CONCLUSION: Using lineage tracing and single cell analytics in mice we show the existence of a unique bone marrow-derived circulating pericyte that homes to ischemia. Our findings have implications in aging, tumorigenesis, and disease.


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