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In vitro models of lymphatic endothelial cells: cytokine receptor expression profiling and incorporation into organotypic culture models

Yang, Fan (2016) In vitro models of lymphatic endothelial cells: cytokine receptor expression profiling and incorporation into organotypic culture models. Master's Thesis, University of Pittsburgh. (Unpublished)

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Abstract

Lymphatic endothelial cells (LECs) line the lymphatic vessels and lymph node sinuses. They function in balancing tissue interstitial fluid, trafficking of dendritic cells and lymphocyte movement into and out of lymph nodes, (lymph node-LECs), and can modulate self-tolerance. LECs also are important for viral pathogenesis and malignant cell migration. The discovery of LEC-specific markers has enabled the isolation and culture of LECs during the past decade, thereby increasing knowledge of LECs biology. Cytokines are small proteins secreted by cells that have pleiotropic effects on cell survival, growth, and functional activities. The expression of cytokine receptors on LECs, if present, could provide insight into the potential functions of LECs. The current study has defined the cytokine receptor expression profile, especially the IL6 family receptors, for three LEC populations: human dermal microvascular lymphatic endothelial cells (HMVEC-dLy), human lung lymphatic microvascular endothelial cells (HMVEC-LLy), and human telomerase reverse transcriptase transfected human dermal lymphatic endothelial cells (hTERT-HDLEC), either with or without viral infection mimicry through the use of poly I:C treatment. Cytokine receptor expression was examined at the RNA, protein and function levels, as a strong reference for further LECs study.
In vitro 3-dimensional (3D) cell culture is an important substitute for in vivo experiments. It is also a meaningful improvement to 2-dimensional cell culture through the modification of culture environments to better mimic in vivo situations. Using an extracellular matrix extraction called MatrigelTM as a simplified 3D model substrate, we demonstrated that five kinds of LECs (HMVEC-dLy, HMVEC-LLy, hTERT-HDLEC, ferret lung isolated LECs and macaque jejunal LECs) can form a network-like structure on it (this process may be regarded as modeling the growth of lymphatic vessels or lymphangiogenesis). In addition, in this study we adapted an organotypic culture model containing lymphatic endothelial cells (LECs) to preliminary build a new powerful tool for in vitro study of LECs and also provide insights into the interactions between LECs and their environment.
The work represented by this thesis holds public health significances lying mainly in enriching the knowledge of human LECs, considering the importance of LECs and lymphatic biology in cancer development and immunology.


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Details

Item Type: University of Pittsburgh ETD
Status: Unpublished
Creators/Authors:
CreatorsEmailPitt UsernameORCID
Yang, Fanfay23@pitt.eduFAY23
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Committee ChairReinhart, Todd A.treinhar@smumn.eduREINHAR
Committee MemberJenkins, Frank J.fjenkins@pitt.eduFJENKINS
Committee MemberChen, Yuecheny@pitt.eduCHENY
Date: 26 March 2016
Date Type: Submission
Defense Date: 1 April 2016
Approval Date: 29 June 2016
Submission Date: 25 April 2016
Access Restriction: 1 year -- Restrict access to University of Pittsburgh for a period of 1 year.
Number of Pages: 73
Institution: University of Pittsburgh
Schools and Programs: Graduate School of Public Health > Infectious Diseases and Microbiology
Degree: MS - Master of Science
Thesis Type: Master's Thesis
Refereed: No
Uncontrolled Keywords: lymphatic endothelial cells; cytokine receptors; IL6; lymphangiogenesis; 3D culture model;
Date Deposited: 29 Jun 2016 19:06
Last Modified: 01 May 2017 05:15
URI: http://d-scholarship.pitt.edu/id/eprint/27449

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