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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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Comparative expression of surface markers in cultured EPC-derived cells, HDLEC and HUVEC by flow cytometric analysis. Cells were labelled with markers for EC, haematopoietic stem cells and monocytes. The black histograms outline the region of fluorescent intensity of the specific antibody and the white histograms that of the negative control antibody. These graphs are representative of five independent experiments.
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fig03: Comparative expression of surface markers in cultured EPC-derived cells, HDLEC and HUVEC by flow cytometric analysis. Cells were labelled with markers for EC, haematopoietic stem cells and monocytes. The black histograms outline the region of fluorescent intensity of the specific antibody and the white histograms that of the negative control antibody. These graphs are representative of five independent experiments.

Mentions: In order to determine the phenotype of EPC-derived cells, we performed flow cytometry and immunohistochemistry analysis using a selected set of markers and compared it to that obtained in HDLEC and HUVEC. The cell surface expression of molecules is summarized in Figure 3 illustrating the major phenotypic differences between the EC types. By FACS analysis, EPC-derived cells were strongly positive for common endothelial markers such as CD31, CD54, CD105 and CD144. These results clearly indicate that the cord blood-derived CD34+ cells had undergone a complete EC differentiation process. In addition, EPC-derived cells were weakly positive for thrombomodulin and CD143 and failed to express CD36. Several cell surface endothelial markers were similarly expressed on HDLEC and HUVEC. Among them were CD31, CD54, CD105 and CD144. Other markers differentiated EPC-derived cells from the other EC types. Unlike EPC-derived cells, HDLEC abundantly expressed CD36, while CD36 was absent on HUVEC. In general, EPC-derived cell cultures did not contain monocytes or macrophages as attested by the absence of CD14+ cells. Differentiation into EC was further associated with the acquisition of HLA-class I antigens, but not HLA-class II antigens. In the line with this observation, EPC-derived cells efficiently expressed β2-microglobulin, which is the light chain of the HLA-class I antigen complex and was much more strongly expressed on HDLEC.


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)

Comparative expression of surface markers in cultured EPC-derived cells, HDLEC and HUVEC by flow cytometric analysis. Cells were labelled with markers for EC, haematopoietic stem cells and monocytes. The black histograms outline the region of fluorescent intensity of the specific antibody and the white histograms that of the negative control antibody. These graphs are representative of five independent experiments.
© Copyright Policy
Related In: Results  -  Collection

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

fig03: Comparative expression of surface markers in cultured EPC-derived cells, HDLEC and HUVEC by flow cytometric analysis. Cells were labelled with markers for EC, haematopoietic stem cells and monocytes. The black histograms outline the region of fluorescent intensity of the specific antibody and the white histograms that of the negative control antibody. These graphs are representative of five independent experiments.
Mentions: In order to determine the phenotype of EPC-derived cells, we performed flow cytometry and immunohistochemistry analysis using a selected set of markers and compared it to that obtained in HDLEC and HUVEC. The cell surface expression of molecules is summarized in Figure 3 illustrating the major phenotypic differences between the EC types. By FACS analysis, EPC-derived cells were strongly positive for common endothelial markers such as CD31, CD54, CD105 and CD144. These results clearly indicate that the cord blood-derived CD34+ cells had undergone a complete EC differentiation process. In addition, EPC-derived cells were weakly positive for thrombomodulin and CD143 and failed to express CD36. Several cell surface endothelial markers were similarly expressed on HDLEC and HUVEC. Among them were CD31, CD54, CD105 and CD144. Other markers differentiated EPC-derived cells from the other EC types. Unlike EPC-derived cells, HDLEC abundantly expressed CD36, while CD36 was absent on HUVEC. In general, EPC-derived cell cultures did not contain monocytes or macrophages as attested by the absence of CD14+ cells. Differentiation into EC was further associated with the acquisition of HLA-class I antigens, but not HLA-class II antigens. In the line with this observation, EPC-derived cells efficiently expressed β2-microglobulin, which is the light chain of the HLA-class I antigen complex and was much more strongly expressed on HDLEC.

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