Transcriptomal comparison of human dermal lymphatic endothelial cells ex vivo and in vitro

Wick, Nikolaus; Saharinen, Pipsa; Saharinen, Juha; Gurnhofer, Elisabeth; Steiner, Carl-Walter; Raab, Ingrid; Stokić, Dejan; Giovanoli, Pietro; Buchsbaum, Sabine; Burchard, Aurea; Thurner, Stefan; Alitalo, Kari; Kerjaschki, Dontscho

doi:10.1152/physiolgenomics.00037.2006

Cited by 95 publications

(96 citation statements)

References 27 publications

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“…Interstitial fluid drains into lymphatic capillaries (also known as initial lymphatics and terminal lymphatics) that are blind-ended vessels with a discontinuous basement membrane that lack pericytes. The interendothelial adhesions are maintained by discontinuous, "button-like" junctions serving to both freely drain interstitial proteins and also facilitate immune cell transmigration [28][29][30][31]. These overlapping cell-cell junctions serve as primary valves to prevent backflow from the lymphatic vessel into the tissue [32] and secondary valves prevent backflow within the vessel [33].…”

Section: Lymphatic Physiology and Neogenesismentioning

confidence: 99%

Role of Lymphatic Vessels in Tumor Immunity: Passive Conduits or Active Participants?

Lund

Swartz

2010

J Mammary Gland Biol Neoplasia

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Research in lymphatic biology and cancer immunology may soon intersect as emerging evidence implicates the lymphatics in the progression of chronic inflammation and autoimmunity as well as in tumor metastasis and immune escape. Like the blood vasculature, the lymphatic system comprises a highly dynamic conduit system that regulates fluid homeostasis, antigen transport and immune cell trafficking, which all play important roles in the progression and resolution of inflammation, autoimmune diseases, and cancer. This review presents emerging evidence that lymphatic vessels are active modulators of immunity, perhaps fine-tuning the response to adjust the balance between peripheral tolerance and immunity. This suggests that the tumor-associated lymphatic vessels and draining lymph node may be important in tumor immunity which in turn governs metastasis.

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Section: Lymphatic Physiology and Neogenesismentioning

confidence: 99%

Role of Lymphatic Vessels in Tumor Immunity: Passive Conduits or Active Participants?

Lund

Swartz

2010

J Mammary Gland Biol Neoplasia

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“…2,3 LEC podo-low and LEC podo-high were purified by fluorescence-activated cell sorting separation of the total LEC fraction by increasing the resolution of the podoplanin channel. The collected cells were lysed for RNA isolation or immobilized on slides by cytospin (Cytospin 3; Dako Cytomation, Glostrup, Denmark) or taken into tissue culture.…”

Section: Isolation Of Endothelial Cellsmentioning

confidence: 99%

“…3,4 The other reagents were FITC-conjugated monoclonal anti-CD31 (catalog no. 555445; BD Biosciences Pharmingen, Franklin Lakes, NJ), RPE-Cy5.1-conjugated mouse monoclonal anti-CD45 (catalog no.…”

Section: Antibodiesmentioning

confidence: 99%

“…The two housekeeping genes in nonquantitative RT-PCR were breakpoint cluster region and ␤ 2-microglobulin, as described previously. 3 In addition, early passage cultures of LEC podo-low and LEC podo-high were subjected to quantitative real-time PCR (Applied Biosystems, Foster City, CA) for relative podoplanin mRNA quantification using ⌬⌬Ct.…”

Section: Rna Analysismentioning

confidence: 99%

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Lymphatic Precollectors Contain a Novel, Specialized Subpopulation of Podoplaninlow, CCL27-Expressing Lymphatic Endothelial Cells

Wick

Haluza

Gurnhofer

et al. 2008

The American Journal of Pathology

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Expression of the lymphoendothelial marker membrane mucoprotein podoplanin (podo) distinguishes endothelial cells of both blood and lymphatic lineages. We have previously discovered two distinct subpopulations of lymphatic endothelial cells (LECs) in human skin that were defined by their cell surface densities of podoplanin and were designated LEC podo-low and LEC -containing precollector vessels constitute a specialized segment of the initial lymphatic microvasculature, and we hypothesize that these LEC podo-low -containing vessels are involved in the trafficking of CCR10 ؉ T cells during skin inflammation.

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“…Cells are often not always and entirely in contact with other cells, as cultures are never grown to 100% confluence. In general, primary cells are also subject to dedifferentiation, and chromosomal instability, characterized by losses or gains of chromosomes during cell replication in continuous cell lines, may be another problem 13,14,15 . The comparison of cultured human fungiform cells with cultured nontaste cells of tongue needs to be explored.…”

Section: Discussionmentioning

confidence: 99%

Isolation and Culture of Human Fungiform Taste Papillae Cells

Ozdener

Spielman

Rawson³

2012

JoVE

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Taste cells are highly specialized, with unique histological, molecular and physiological characteristics that permit detection of a wide range of simple stimuli and complex chemical molecules contained in foods. In human, individual fungiform papillae contain from zero to as many as 20 taste buds. There is no established protocol for culturing human taste cells, although the ability to maintain taste papillae cells in culture for multiple cell cycles would be of considerable utility for characterizing the molecular, regenerative, and functional properties of these unique sensory cells. Earlier studies of taste cells have been done using freshly isolated cells in primary culture, explant cultures from rodents, or semi-intact taste buds in tissue slices 1,2,3,4 . Although each of these preparations has advantages, the development of long-term cultures would have provided significant benefits, particularly for studies of taste cell proliferation and differentiation. Several groups, including ours, have been interested in the development and establishment of taste cell culture models. Most attempts to culture taste cells have reported limited viability, with cells typically not lasting beyond 3-5 d 5,6,7,8 . We recently reported on a successful method for the extended culture of rodent taste cells 9 . We here report for the first time the establishment of an in vitro culture system for isolated human fungiform taste papillae cells. Cells from human fungiform papillae obtained by biopsy were successfully maintained in culture for more than eight passages (12 months) without loss of viability. Cells displayed many molecular and physiological features characteristic of mature taste cells. Gustducin and phospholipase C β2, (PLC-β2) mRNA were detected in many cells by reverse transcriptase-polymerase chain reaction and confirmed by sequencing. Immunocytochemistry analysis demonstrated the presence of gustducin and PLC-β2 expression in cultured taste cells. Cultured human fungiform cells also exhibited increases in intracellular calcium in response to appropriate concentrations of several taste stimuli indicating that taste receptors and at least some of the signalling pathways were present. These results sufficient indicate that taste cells from adult humans can be generated and maintained for at least eight passages. Many of the cells retain physiological and biochemical characteristics of acutely isolated cells from the adult taste epithelium to support their use as a model taste system. This system will enable further studies of the processes involved in proliferation, differentiation and function of mammalian taste receptor cells in an in vitro preparation.Human fungiform taste papillae used for establishing human fungiform cell culture were donated for research following proper informed consent under research protocols that were reviewed and approved by the IRB committee. The protocol (#0934) was approved by Schulman Associates Institutional Review Board Inc., Cincinnati, OH. Written protocol below is based...

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Transcriptomal comparison of human dermal lymphatic endothelial cells ex vivo and in vitro

Cited by 95 publications

References 27 publications

Role of Lymphatic Vessels in Tumor Immunity: Passive Conduits or Active Participants?

Role of Lymphatic Vessels in Tumor Immunity: Passive Conduits or Active Participants?

Lymphatic Precollectors Contain a Novel, Specialized Subpopulation of Podoplaninlow, CCL27-Expressing Lymphatic Endothelial Cells

Isolation and Culture of Human Fungiform Taste Papillae Cells

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