eprintid: 27449
rev_number: 37
userid: 5581
dir: disk0/00/02/74/49
datestamp: 2016-06-29 19:06:51
lastmod: 2017-05-01 05:15:11
status_changed: 2016-06-29 19:06:51
type: thesis_degree
metadata_visibility: show
contact_email: fyang2003@outlook.com
item_issues_count: 0
eprint_status: archive
creators_name: Yang, Fan
creators_email: fay23@pitt.edu
creators_id: FAY23
title: In vitro models of lymphatic endothelial cells:  cytokine receptor expression profiling and incorporation into organotypic culture models
ispublished: unpub
divisions: sch_gsph_infectiousdiseasesmicrobiology
full_text_status: public
keywords: lymphatic endothelial cells; cytokine receptors; IL6; lymphangiogenesis; 3D culture model;
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.
date: 2016-03-26
date_type: submitted
pages: 73
institution: University of Pittsburgh
refereed: FALSE
etdcommittee_type: committee_chair
etdcommittee_type: committee_member
etdcommittee_type: committee_member
etdcommittee_name: Reinhart, Todd A.
etdcommittee_name: Jenkins, Frank J.
etdcommittee_name: Chen, Yue
etdcommittee_email: treinhar@smumn.edu
etdcommittee_email: fjenkins@pitt.edu
etdcommittee_email: cheny@pitt.edu
etdcommittee_id: REINHAR
etdcommittee_id: FJENKINS
etdcommittee_id: CHENY
etd_defense_date: 2016-04-01
etd_approval_date: 2016-06-29
etd_submission_date: 2016-04-25
etd_release_date: 2016-06-29
etd_access_restriction: 1_year
etd_patent_pending: FALSE
thesis_type: thesis
degree: MS
citation:   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)  
document_url: http://d-scholarship-dev.library.pitt.edu/27449/1/Fan_Yang_MS_thesis_4_2016.pdf