Isolation and Characterization of Dermal Lymphatic and Blood Endothelial Cells Reveal Stable and Functionally Specialized Cell Lineages

Kriehuber, Ernst; Breiteneder-Geleff, Silvana; Groeger, Marion; Soleiman, Afschin; Schoppmann, Sebastian F.; Stingl, Georg; Kerjaschki, Dontscho; Maurer, Dieter

doi:10.1084/jem.194.6.797

Cited by 454 publications

(466 citation statements)

References 39 publications

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“…Podoplanin is a 43 kD cell surface glycoprotein expressed on lymphatic but not blood endothelia (Kriehuber et al 2001). Together with vascular endothelial growth factor receptor 3 (VEGFR-3) and the hyaluronan receptor LYVE-1, podoplanin constitute an important marker for lymphatic vessels, e.g.…”

Section: Introductionmentioning

confidence: 99%

An Interstitial Network of Podoplanin-Expressing Cells in the Human Endolymphatic Duct

Ekwall

Mayerl

Rubin

et al. 2006

JARO

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The human endolymphatic duct (ED) with encompassing interstitial connective tissue (CT) is believed to be important for endolymph resorption and fluid pressure regulation of the inner ear. The periductal CT cells are interconnected via numerous cellular extensions, but do not form vessel structures. Here we report that the periductal CT is populated by two distinct cell phenotypes; one expressing podoplanin, a protein otherwise found on lymph endothelia and on epithelia involved in fluid fluxes, and a second expressing a fibroblast marker. A majority of the interstitial cells expressed podoplanin but not the lymphatic endothelial cell markers hyaluronan receptor (LYVE-1) or vascular endothelial growth factor receptor-3 (VEGFR-3). The fibroblast marker positive cells were found close to the ED epithelium. In the mid-and distal parts of the ED, these cells were enriched under folded epithelia. Furthermore, subepithelial CT cells were found to express activated platelet derived growth factor (PDGF)-" receptors. Cultured CT cells from human inner ear periductal and perisaccular explant tissues were identified as fibroblasts. These cells compacted a three-dimensional collagen lattice by a process that could be promoted by PDGF-BB, a factor involved in interstitial fluid pressure regulation. Our results are compatible with the notion that the periductal CT cells are involved in the regulation of inner ear fluid pressure. By active compaction of the periductal CT and by the formation of villous structures, the CT cells could modulate fluid fluxes over the ED epithelium as well as the longitudinal flow of endolymph in the ED.

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Section: Introductionmentioning

confidence: 99%

An Interstitial Network of Podoplanin-Expressing Cells in the Human Endolymphatic Duct

Ekwall

Mayerl

Rubin

et al. 2006

JARO

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“…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 receptor-3 (VEGFR-3), a transmembrane tyrosine kinase receptor for vascular endothelial growth factors-C (VEGF-C) and VEGF-D. 4,5 Recent reports indicate that the expression of the integrin ␣9 subunit is restricted to lymphatic endothelium. 6 Use of these markers has permitted the isolation of relatively pure populations of BECs and LECs using fluorescence-activated cell sorting (FACS) 7 or immunomag-netic beads. 8,9 Initial characterization of these populations highlighted major differences between the two cell types.…”

mentioning

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

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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...

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“…Major advances in lymphatic research have been enabled by the recent establishment of defined cultures of blood vascular and lymphatic endothelial cells isolated from human skin (Kriehuber et al, 2001;Makinen et al, 2001b;Podgrabinska et al, 2002;Hirakawa et al, 2003). These cells maintain their lineage-specific differentiation even after several passages in vitro.…”

Section: Molecular Control Of Lymphatic Development and Function In Mmentioning

confidence: 99%

“…VEGFR-2, also known as KDR or Flk1, is a receptor for VEGF-A, VEGF-C, and VEGF-D, and is expressed by blood vascular and lymphatic endothelial cells in situ and in vitro (Kriehuber et al, 2001;Hirakawa et al, 2003;. Although the role of VEGFR-2 in angiogenesis has been well established, the possible contribution of VEGF-A signaling through VEGFR-2 to lymphangiogenesis is currently a matter of controversy.…”

Section: Vegfr-2mentioning

confidence: 99%

“…Podoplanin is a mucin-type transmembrane glycoprotein that is expressed by lymphatic, but not by blood vascular endothelial cells in vivo and in vitro (Wetterwald et al, 1996;Kriehuber et al, 2001;Petrova et al, 2002;Hirakawa et al, 2003;Schacht et al, 2003). In addition, podoplanin is expressed by several other cell types, including type I alveolar cells of the lung, cells of the choroid plexus, ciliary epithelial cells of the eye, osteocytes and kidney podocytes, and multiple synonyms have been used in the literature, such as RTI40, murine gp38, canine gp40, human gp36, and murine PA2.26 Schacht et al, 2003).…”

Section: Podoplaninmentioning

confidence: 99%

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Development of the lymphatic vascular system: A mystery unravels

Hong

Shin

Detmar

2004

Developmental Dynamics

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The blood vascular and the lymphatic system play complementary roles in tissue perfusion and fluid reabsorption. Despite its critical role in mediating tissue fluid homeostasis, intestinal lipid absorption, and the immune response, the lymphatic system has not received as much attention as the blood vascular system, largely due to a lack of lymphatic-specific markers and to the dearth of knowledge about the molecular regulation of lymphatic development and function. A series of recent landmark studies now significantly has advanced our understanding of the lymphatic system. Based upon the discovery and characterization of lymphatic-specific growth factors, receptors, and transcriptional regulators, the mystery of lymphatic vascular system development begins to be unraveled. The successful isolation and cultivation of blood vascular and lymphatic endothelial cells has enabled comparative molecular and cellular analyses of these two genetically and developmentally closely related cell lineages. Moreover, studies of several genetic mouse models have set the framework for a new molecular model of embryonic lymphatic vascular development and have identified molecular pathways whose mutational inactivation leads to human diseases associated with lymphedema. Although these rapid advances already have led to development of the first lymphatic-targeted molecular therapies, there still remain many unanswered questions regarding almost every aspect of lymphatic vascular biology, making the lymphatic system a highly exciting and rewarding field of study.

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Isolation and Characterization of Dermal Lymphatic and Blood Endothelial Cells Reveal Stable and Functionally Specialized Cell Lineages

Cited by 454 publications

References 39 publications

An Interstitial Network of Podoplanin-Expressing Cells in the Human Endolymphatic Duct

An Interstitial Network of Podoplanin-Expressing Cells in the Human Endolymphatic Duct

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

Development of the lymphatic vascular system: A mystery unravels

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