Telomerized human microvasculature is functional in vivo

Yang, Jiwei; Nagavarapu, Usha; Relloma, Kenneth; Sjaastad, Michael D.; Moss, William C.; Passaniti, Antonino; Herron, G. Scott

doi:10.1038/85655

Cited by 138 publications

(100 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…estingly, hTERT-immortalized human microvascular endothelial cells have been reported to form functional capillaries in vivo. 47 However, blood vascular versus lymphatic phenotypic characterization was not assessed in these cells.…”

Section: Discussionmentioning

confidence: 99%

Generation and Characterization of Telomerase-Transfected Human Lymphatic Endothelial Cells with an Extended Life Span

Nisato

Buser

et al. 2004

The American Journal of Pathology

View full text Add to dashboard Cite

The study of lymphatic endothelial cells and lymphangiogenesis has, in the past, been hampered by the lack of lymphatic endothelial-specific markers. The recent discovery of several such markers has permitted the isolation of lymphatic endothelial cells (LECs) from human skin. However, cell numbers are limited and purity is variable with the different isolation procedures. To overcome these problems, we have transfected human dermal microvascular endothelial cells (HDMVECs) with a retrovirus containing the coding region of human telomerase reverse transcriptase (hTERT), and have produced a cell line, hTERT-HDLEC, with an extended lifespan. hTERT-HDLEC exhibit a typical cobblestone morphology when grown in culture, are contact-inhibited, and express endothelial cell-specific markers. hTERT-HDLEC also express the recognized lymphatic markers, Prox-1, LYVE-1 and podoplanin, as well as integrin ␣9, but do not express CD34. They also form tube-like structures in three-dimensional collagen gels when stimulated with vascular endothelial growth factors -A and -C. Based on these currently recognized criteria, these cells are LEC. Surprisingly, we also found that the widely studied HMEC-1 cell line expresses recognized lymphatic markers; however, these cells are also CD34-positive. In summary, the ectopic expression of hTERT increases the life span of LECs and does not affect their capacity to form tubelike structures in a collagen matrix. The production and characterization of hTERT-HDLEC will facilitate the study of the properties of lymphatic endothelium in vitro. The blood and lymphatic vascular systems are anatomically and histologically closely related but distinct, and do not anastomose except at the level of the main lymphatic ducts. The blood vascular system supplies nutrients and oxygen to the body while the lymphatic vasculature is crucial for immune cell traffic and the uptake of extracellular fluid. The formation of new blood vessels from pre-existing blood vessels (angiogenesis) occurs in physiological (eg, postnatal growth, inflammation, and tissue repair) and pathological (eg, tumor recruitment of blood vessels) settings. Lymphangiogenesis is the formation of new lymphatic vessels from pre-existing lymphatic capillaries, and occurs in similar settings. Blood endothelial cells (BECs) and lymphatic endothelial cells (LECs) are quiescent cells that line the lumina of blood and lymphatic vessels.The lymphatic system has traditionally been overshadowed by the greater emphasis placed on the blood vascular system. This has been due in part to the absence of suitable markers that distinguish lymphatic from blood vascular endothelium. In the past few years, this limitation has been overcome. Lymphatic markers include LYVE-1, a lymphatic endothelial receptor for the extracellular matrix/lymphatic fluid glycosaminoglycan hyaluronan; 1 Prox-1, a homeobox gene product involved in regulating early lymphatic development; 2 podoplanin, a glomerular podocyte membrane mucoprotein; 3 and vascular endothelial growth factor re...

show abstract

Section: Discussionmentioning

confidence: 99%

Generation and Characterization of Telomerase-Transfected Human Lymphatic Endothelial Cells with an Extended Life Span

Nisato

Buser

et al. 2004

The American Journal of Pathology

View full text Add to dashboard Cite

show abstract

“…The second restraint is much more important because it determines the overall feasibility of the technique, whereas apoptosis can be prevented either by using telomerized cells or cells transfected with blc-2 (Yang et al, 2001;Nor et al, 2001). Based on estimations of the optimum minimal thickness, the diameter of the cell aggregates must be no smaller than 600 m. This does not exclude the possibility of variability in cell aggregate size, but it definitely places a well-defined limit on the smallest technologically acceptable diameter.…”

Section: Identification Of the Optimum Size For Cell Aggregatesmentioning

confidence: 99%

Cell and organ printing 2: Fusion of cell aggregates in three‐dimensional gels

et al. 2003

View full text Add to dashboard Cite

We recently developed a cell printer (Wilson and Boland, 2003) that enables us to place cells in positions that mimic their respective positions in organs. However, this technology was limited to the printing of two-dimensional (2D) tissue constructs. Here we describe the use of thermosensitive gels to generate sequential layers for cell printing. The ability to drop cells on previously printed successive layers provides a real opportunity for the realization of three-dimensional (3D) organ printing. Organ printing will allow us to print complex 3D organs with computer-controlled, exact placing of different cell types, by a process that can be completed in several minutes. To demonstrate the feasibility of this novel technology, we showed that cell aggregates can be placed in the sequential layers of 3D gels close enough for fusion to occur. We estimated the optimum minimal thickness of the gel that can be reproducibly generated by dropping the liquid at room temperature onto a heated substrate. Then we generated cell aggregates with the corresponding (to the minimal thickness of the gel) size to ensure a direct contact between printed cell aggregates during sequential printing cycles. Finally, we demonstrated that these closely-placed cell aggregates could fuse in two types of thermosensitive 3D gels. Taken together, these data strongly support the feasibility of the proposed novel organ-printing technology. Anat Rec Part A 272A: 497-502, 2003.

show abstract

“…Approximately 1 ϫ 10 6 human dermal microvascular endothelial cells were mixed with 0.5 ml of Matrigel on ice in the presence of either antibody 2A3 or control IgM, and the mixture was implanted into the ventral midline thoracic tissue of a mouse following procedures outlined in ref. 24. At 7 days, the implants were removed, fixed and processed for immunofluorescence microscopy as above.…”

Section: Methodsmentioning

confidence: 99%

Complex interactions between the laminin α4 subunit and integrins regulate endothelial cell behavior in vitro and angiogenesis in vivo

Gonzalez

Gonzales

Herron

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

114

View full text Add to dashboard Cite

The ␣4 laminin subunit is a component of the basement membrane of blood vessels where it codistributes with the integrins ␣v␤3, ␣3␤1, and ␣6␤1. An antibody against the G domain (residues 919-1207; G 919 -1207 ) of the ␣4 laminin subunit inhibits angiogenesis in a mouse-human chimeric model, indicating the functional importance of this domain. Additional support for the latter derives from the ability of recombinant G 919 -1207 to support endothelial cell adhesion. In particular, endothelial cell adhesion to G 919 -1207 is half-maximal at 1.4 nM, whereas residues 919-1018 and 1016 -1207 of the G domain are poor cellular ligands. Function blocking antibodies against integrins ␣v␤3 and ␤1 and a combination of antibodies against ␣3 and ␣6 integrin subunits inhibit endothelial cell attachment to G 919 -1207 . Moreover, both ␣v␤3 and ␣3␤1 integrin bind with high affinity to G 919 -1207 . Together, our studies demonstrate that the G domain of laminin ␣4 chain is a specific, high affinity ligand for the ␣v␤3 and ␣3␤1 integrin heterodimers and that these integrins, together with ␣6␤1, function cooperatively to mediate endothelial cell-␣4 laminin interaction and hence blood vessel development. We propose a model based on these data that reconcile apparent discrepancies in the recent literature with regard to the role of the ␣v␤3 integrin in angiogenesis. matrix ͉ matrix receptor ͉ blood vessels

show abstract

Telomerized human microvasculature is functional in vivo

Cited by 138 publications

References 36 publications

Generation and Characterization of Telomerase-Transfected Human Lymphatic Endothelial Cells with an Extended Life Span

Generation and Characterization of Telomerase-Transfected Human Lymphatic Endothelial Cells with an Extended Life Span

Cell and organ printing 2: Fusion of cell aggregates in three‐dimensional gels

Complex interactions between the laminin α4 subunit and integrins regulate endothelial cell behavior in vitro and angiogenesis in vivo

Contact Info

Product

Resources

About