Mouse lung contains endothelial progenitors with high capacity to form blood and lymphatic vessels

Abstract

BackgroundPostnatal endothelial progenitor cells (EPCs) have been successfully isolated from whole bone marrow, blood and the walls of conduit vessels. They can, therefore, be classified into circulating and resident progenitor cells. The differentiation capacity of resident lung endothelial progenitor cells from mouse has not been evaluated.ResultsIn an attempt to isolate differentiated mature endothelial cells from mouse lung we found that the lung contains EPCs with a high vasculogenic capacity and capability of de novo vasculogenesis for blood and lymph vessels.Mouse lung microvascular endothelial cells (MLMVECs) were isolated by selection of CD31+ cells. Whereas the majority of the CD31+ cells did not divide, some scattered cells started to proliferate giving rise to large colonies (> 3000 cells/colony). These highly dividing cells possess the capacity to integrate into various types of vessels including blood and lymph vessels unveiling the existence of local microvascular endothelial progenitor cells (LMEPCs) in adult mouse lung. EPCs could be amplified > passage 30 and still expressed panendothelial markers as well as the progenitor cell antigens, but not antigens for immune cells and hematopoietic stem cells. A high percentage of these cells are also positive for Lyve1, Prox1, podoplanin and VEGFR-3 indicating that a considerabe fraction of the cells are committed to develop lymphatic endothelium. Clonogenic highly proliferating cells from limiting dilution assays were also bipotent. Combined in vitro and in vivo spheroid and matrigel assays revealed that these EPCs exhibit vasculogenic capacity by forming functional blood and lymph vessels.ConclusionThe lung contains large numbers of EPCs that display commitment for both types of vessels, suggesting that lung blood and lymphatic endothelial cells are derived from a single progenitor cell.

The isolation and culture of endothelial colony-forming cells from human and rat lungs

“…Recently, three papers have demonstrated the existence of endothelial progenitor-like cells with a high potential for proliferation in the peripheral microvasculature of rat 19,20 and mouse 21 lungs.…”

“…The cells were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) and antibiotics and observed to demonstrate ECFC-like phenotype, on the basis of both expression of surface antigens and functional behaviours such as uptake of low-density lipoprotein (LDL), maintenance of high barrier resistance, in vitro and in vivo vasculogenesis in Matrigel 20 . Weich and colleagues 21,22 , on the other hand, selected out CD31+ cells from mouse peripheral lung microvasculature by successively using magnetic and fluorescence-activated cell sorting (MACS and FACS). They expanded these cells in DMEM enriched with 20% FBS and demonstrate their expression of panendothelial cell surface markers 22 and ability to bind Griffonia simplicifolia lectin.…”

Cortactin deficiency is associated with reduced neutrophil recruitment but increased vascular permeability in vivo

“…Murine bone marrowderived leukocytes were isolated as described previously (Bixel et al, 2007). Primary MLECs were isolated from 8-wk-old Cttn del/del and littermate Cttn WT/WT mice as described previously (Schniedermann et al, 2010).…”

Fusing VE-Cadherin to α-Catenin Impairs Fetal Liver Hematopoiesis and Lymph but Not Blood Vessel Formation

“…Primary endothelial cells were isolated from lungs of 7-to 9-week-old Cdh5 wt/wt or Cdh5 VEC␣C/VEC␣C mice as previously described (26). CD31-positive endothelial cells were additionally sorted by flow cytometry for lymphatic identity using a PE coupled podoplanin antibody (BD Biosciences).…”