VEGFR2+PDGFRβ+ circulating precursor cells participate in capillary restoration after hyperoxia acute lung injury (HALI)
Corresponding Author
Rosemary Jones
Department of Anesthesia and Critical Care, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA
Correspondence to: Rosemary C. JONES, Ph.D., CNY-3416, Building 149, 13th Street, Charlestown, MA 02129, USA.Tel.: (617) 726-4359Fax: (617) 726-4374E-mail: [email protected]Dan G. DUDA, D.M.D., Ph.D., CNY-3402, Building 149, 13th Street, Charlestown, MA 02129, USA.Tel.: (617) 726-4648Fax: (617) 726-1962E-mail: [email protected]Search for more papers by this authorDiane E. Capen
Department of Anesthesia and Critical Care, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA
Search for more papers by this authorMargaretha Jacobson
Department of Anesthesia and Critical Care, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA
Search for more papers by this authorKenneth S. Cohen
Center for Regenerative Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA
Current affiliation: Section of Hematology/Oncology, Department of Medicine, University of Chicago, Chicago, IL.
Search for more papers by this authorDavid T. Scadden
Center for Regenerative Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA
Search for more papers by this authorCorresponding Author
Dan G. Duda
Steele Laboratory, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA
Correspondence to: Rosemary C. JONES, Ph.D., CNY-3416, Building 149, 13th Street, Charlestown, MA 02129, USA.Tel.: (617) 726-4359Fax: (617) 726-4374E-mail: [email protected]Dan G. DUDA, D.M.D., Ph.D., CNY-3402, Building 149, 13th Street, Charlestown, MA 02129, USA.Tel.: (617) 726-4648Fax: (617) 726-1962E-mail: [email protected]Search for more papers by this authorCorresponding Author
Rosemary Jones
Department of Anesthesia and Critical Care, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA
Correspondence to: Rosemary C. JONES, Ph.D., CNY-3416, Building 149, 13th Street, Charlestown, MA 02129, USA.Tel.: (617) 726-4359Fax: (617) 726-4374E-mail: [email protected]Dan G. DUDA, D.M.D., Ph.D., CNY-3402, Building 149, 13th Street, Charlestown, MA 02129, USA.Tel.: (617) 726-4648Fax: (617) 726-1962E-mail: [email protected]Search for more papers by this authorDiane E. Capen
Department of Anesthesia and Critical Care, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA
Search for more papers by this authorMargaretha Jacobson
Department of Anesthesia and Critical Care, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA
Search for more papers by this authorKenneth S. Cohen
Center for Regenerative Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA
Current affiliation: Section of Hematology/Oncology, Department of Medicine, University of Chicago, Chicago, IL.
Search for more papers by this authorDavid T. Scadden
Center for Regenerative Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA
Search for more papers by this authorCorresponding Author
Dan G. Duda
Steele Laboratory, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA
Correspondence to: Rosemary C. JONES, Ph.D., CNY-3416, Building 149, 13th Street, Charlestown, MA 02129, USA.Tel.: (617) 726-4359Fax: (617) 726-4374E-mail: [email protected]Dan G. DUDA, D.M.D., Ph.D., CNY-3402, Building 149, 13th Street, Charlestown, MA 02129, USA.Tel.: (617) 726-4648Fax: (617) 726-1962E-mail: [email protected]Search for more papers by this authorAbstract
The in vivo morphology and phenotype of circulating cells that spontaneously contribute to new vessel formation in adults remain unclear. Here, we use high-resolution imaging and flow cytometry to characterize the morphology and phenotype of a distinct population of circulating mononuclear cells contributing to spontaneous new vessel formation after hyperoxia acute lung injury (HALI). We identify a subpopulation of myeloid (CD11b/Mac1+) haematopoietic cells co-expressing vascular endothelial growth factor receptor 2 (VEGFR2) and platelet derived growth factor receptor beta (PDGFRβ). Moreover, we show that these CD11b+VEGFR2+PDGFRβ+ circulating precursor cells (CPCs) contribute structurally to the luminal surface of capillaries re-forming 2 weeks post-HALI. This indicates that these myeloid CPCs may function, at least transiently, as putative vascular precursors, and has important implications for capillary growth and repair in injury and in pathologies of the lung and other organs.
Supporting Information
Fig. S1 CPC morphology is distinct from that of mature hematopoietic cells circulating in lung capillaries. Representative image of CPC for comparison with a circulating PMN, eosinophil, mast cell and monocyte. Each cell type is clearly identified by its characteristic arrangement of nuclear heterochromatin and cytoplasmic organelles. A dense arrangement of free ribosomes and paucity of organelles other than mitochrondria characterize the cytosol of the CPC (a and b); in addition to a highly lobulated nucleus, an electron-dense cytosol with numerous granules characterize the PMN (c and d); a lobulated nucleus and granules with a crystalline structure characterize the eosinophil (e); the monocyte is characterized by a typical horseshoe-shaped nucleus and a sparse arrangement of nuclear heterochomatin to euchromatin (f). 80 nm-thick epon resin sections stained with uranyl acetate and lead citrate. Bars = 1 mm (a-f).
Fig. S2 CPCs contact endothelium post-HALI. Representative image of CPC forming small contact sites (arrows) with the process of an adjacent endothelial cell post-HALI (week 6). Note the large mitochondria (MT) and dense arrangement of free ribsomes in the CPC cytosol. 80 nm-thick epon resin section stained with uranyl acetate and lead citrate: Bar = 0.5 mm.
Fig. S3 CPCs align as capillary perivascular cells post-HALI. Region of the alveolar capillary membrane (post-HALI, week 7), with multiple capillary structures (evident by erythrocytes in their lumen) and with CPCs that have crossed a capillary wall into the interstitium (a, see boxed area). At higher magnification, these cells are seen to retain the typical morphology of CPCs within the capillary lumen: note the extended process of the cell aligning as a perivascular cell along the abluminal surface of a capillary wall (b, arrows). High magnification illustrating 10 nm protein-A gold labeled CD11b antigenic sites (inset) typically expressed by capillary perivascular cells in the model (post-HALI, week 6). Red delineates the plasmalemmal membrane of CPCs; blue delineates that of capillary endothelial cell processes. 80 nm-thick epon resin sections stained with uranyl acetate and lead citrate. Bars = 10 mm (a), 6 mm (b), 0.1 mm (c).
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