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Endothelial cells from cord blood CD133+CD34+ progenitors share phenotypic, functional and gene expression profile similarities with lymphatics.

Nguyen VA, Fürhapter C, Obexer P, Stössel H, Romani N, Sepp N - J. Cell. Mol. Med. (2009)

Bottom Line: Structures compatible with Weibel-Palade bodies were also found by electron microscopy.Among them were adhesion molecules, extracellular matrix proteins and cytokines.Our data point to the close lineage relationship of both types of vascular cells and support the theory of a venous origin of the lymphatic system.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, Innsbruck Medical University, Innsbruck, Austria. van.nguyen@i-med.ac.at

ABSTRACT
The existence of endothelial progenitor cells (EPC) with high cell-cycle rate in human umbilical cord blood has been recently shown and represents a challenging strategy for therapeutic neovascularization. To enhance knowledge for future cellular therapy, we compared the phenotypic, functional and gene expression differences between EPC-derived cells generated from cord blood CD34(+) cells, and lymphatic and macrovascular endothelial cells (EC) isolated from human foreskins and umbilical veins, respectively. Under appropriate culture conditions, EPC developed into fully matured EC with expression of similar endothelial markers as lymphatic and macrovascular EC, including CD31, CD36, von Willebrand factor FVIII, CD54 (ICAM-1), CD105 (endoglin), CD144 (VE-cadherin), Tie-1, Tie-2, VEGFR-1/Flt-1 and VEGFR-2/Flk-1. Few EPC-derived cells became positive for LYVE-1, indicating their origin from haematopoietic stem cells. However they lacked expression of other lymphatic cell-specific markers such as podoplanin and Prox-1. Functional tests demonstrated that the cobblestone EPC-derived cells up-regulated CD54 and CD62E expression in response to TNF-alpha, incorporated DiI-acetylated low-density liproprotein and formed cord- and tubular-like structures with capillary lumen in three-dimensional collagen culture--all characteristic features of the vascular endothelium. Structures compatible with Weibel-Palade bodies were also found by electron microscopy. Gene microarray profiling revealed that only a small percentage of genes investigated showed differential expression in EPC-derived cells and lymphatic EC. Among them were adhesion molecules, extracellular matrix proteins and cytokines. Our data point to the close lineage relationship of both types of vascular cells and support the theory of a venous origin of the lymphatic system.

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Weibel-Palade bodies in cord blood CD34+-derived EPC-derived cells. (B) and (D) show Weibel-Palade bodies in the cell that is depicted at low power in (A). Note the typical internal structure of the bodies. For comparison, (C) shows a Weibel-Palade body from a dermal EC in healthy human skin in situ. (B-D) are at the same magnification. Final magnifications: (A) ×6.200, (B-D) ×90.000; scale bars correspond to 2 μm in (A) and 100 nm in (B-D).
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fig02: Weibel-Palade bodies in cord blood CD34+-derived EPC-derived cells. (B) and (D) show Weibel-Palade bodies in the cell that is depicted at low power in (A). Note the typical internal structure of the bodies. For comparison, (C) shows a Weibel-Palade body from a dermal EC in healthy human skin in situ. (B-D) are at the same magnification. Final magnifications: (A) ×6.200, (B-D) ×90.000; scale bars correspond to 2 μm in (A) and 100 nm in (B-D).

Mentions: Ultrastructurally, cells of variable sizes were found; some cells were extraordinarily large. They were often polarized in that a dense network of villi protruded from one pole of the cell. At the basis of these villi, micropinocytic vesicles could be detected. Sometimes they were abundant, thus resembling dermal EC in situ, but they were only found there in substantial numbers. The cells possessed large nuclei that were deeply indented one- or severalfold, similar to EC in situ, and contained many autophagic vacuoles filled with little membrane vesicles (reminiscent of mul-tivesicular bodies) and often large membrane whorls (myelinoid figures). Lipid droplets occasionally occurred, presumably a consequence of the long culture period. The cells were rich in ribosomes, both as free ribosomes and as ribosomes attached to the endoplasmic reticulum. Rarely, some stretches of rough endoplasmic reticulum were arranged in a parallel fashion, resembling ‘ergastoplasma’ as observed in plasma cells. Intermediate filaments were easily detected. Only rarely, they were arranged in the parallel, ‘spaghetti-like’ fashion as known from EC in situ. Unequivocal Weibel-Palade bodies were scarce, scarcer as compared to EC in situ. They could only be spotted in a fraction of all the section profiles (less than one in ten) (Fig. 2).


Endothelial cells from cord blood CD133+CD34+ progenitors share phenotypic, functional and gene expression profile similarities with lymphatics.

Nguyen VA, Fürhapter C, Obexer P, Stössel H, Romani N, Sepp N - J. Cell. Mol. Med. (2009)

Weibel-Palade bodies in cord blood CD34+-derived EPC-derived cells. (B) and (D) show Weibel-Palade bodies in the cell that is depicted at low power in (A). Note the typical internal structure of the bodies. For comparison, (C) shows a Weibel-Palade body from a dermal EC in healthy human skin in situ. (B-D) are at the same magnification. Final magnifications: (A) ×6.200, (B-D) ×90.000; scale bars correspond to 2 μm in (A) and 100 nm in (B-D).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3822512&req=5

fig02: Weibel-Palade bodies in cord blood CD34+-derived EPC-derived cells. (B) and (D) show Weibel-Palade bodies in the cell that is depicted at low power in (A). Note the typical internal structure of the bodies. For comparison, (C) shows a Weibel-Palade body from a dermal EC in healthy human skin in situ. (B-D) are at the same magnification. Final magnifications: (A) ×6.200, (B-D) ×90.000; scale bars correspond to 2 μm in (A) and 100 nm in (B-D).
Mentions: Ultrastructurally, cells of variable sizes were found; some cells were extraordinarily large. They were often polarized in that a dense network of villi protruded from one pole of the cell. At the basis of these villi, micropinocytic vesicles could be detected. Sometimes they were abundant, thus resembling dermal EC in situ, but they were only found there in substantial numbers. The cells possessed large nuclei that were deeply indented one- or severalfold, similar to EC in situ, and contained many autophagic vacuoles filled with little membrane vesicles (reminiscent of mul-tivesicular bodies) and often large membrane whorls (myelinoid figures). Lipid droplets occasionally occurred, presumably a consequence of the long culture period. The cells were rich in ribosomes, both as free ribosomes and as ribosomes attached to the endoplasmic reticulum. Rarely, some stretches of rough endoplasmic reticulum were arranged in a parallel fashion, resembling ‘ergastoplasma’ as observed in plasma cells. Intermediate filaments were easily detected. Only rarely, they were arranged in the parallel, ‘spaghetti-like’ fashion as known from EC in situ. Unequivocal Weibel-Palade bodies were scarce, scarcer as compared to EC in situ. They could only be spotted in a fraction of all the section profiles (less than one in ten) (Fig. 2).

Bottom Line: Structures compatible with Weibel-Palade bodies were also found by electron microscopy.Among them were adhesion molecules, extracellular matrix proteins and cytokines.Our data point to the close lineage relationship of both types of vascular cells and support the theory of a venous origin of the lymphatic system.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, Innsbruck Medical University, Innsbruck, Austria. van.nguyen@i-med.ac.at

ABSTRACT
The existence of endothelial progenitor cells (EPC) with high cell-cycle rate in human umbilical cord blood has been recently shown and represents a challenging strategy for therapeutic neovascularization. To enhance knowledge for future cellular therapy, we compared the phenotypic, functional and gene expression differences between EPC-derived cells generated from cord blood CD34(+) cells, and lymphatic and macrovascular endothelial cells (EC) isolated from human foreskins and umbilical veins, respectively. Under appropriate culture conditions, EPC developed into fully matured EC with expression of similar endothelial markers as lymphatic and macrovascular EC, including CD31, CD36, von Willebrand factor FVIII, CD54 (ICAM-1), CD105 (endoglin), CD144 (VE-cadherin), Tie-1, Tie-2, VEGFR-1/Flt-1 and VEGFR-2/Flk-1. Few EPC-derived cells became positive for LYVE-1, indicating their origin from haematopoietic stem cells. However they lacked expression of other lymphatic cell-specific markers such as podoplanin and Prox-1. Functional tests demonstrated that the cobblestone EPC-derived cells up-regulated CD54 and CD62E expression in response to TNF-alpha, incorporated DiI-acetylated low-density liproprotein and formed cord- and tubular-like structures with capillary lumen in three-dimensional collagen culture--all characteristic features of the vascular endothelium. Structures compatible with Weibel-Palade bodies were also found by electron microscopy. Gene microarray profiling revealed that only a small percentage of genes investigated showed differential expression in EPC-derived cells and lymphatic EC. Among them were adhesion molecules, extracellular matrix proteins and cytokines. Our data point to the close lineage relationship of both types of vascular cells and support the theory of a venous origin of the lymphatic system.

Show MeSH
Related in: MedlinePlus