Isolation and characterization of lymphatic endothelial cells isolated from rat mesentery : a thesis /
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| Format: | Thesis Book |
| Language: | English |
| Published: |
[College Station, Tex.] :
[Texas A&M University System Health Science Center],
[2005]
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| Subjects: |
| Abstract: | ABSTRACT: The lymphatic endothelium is important to the functioning of the lymphatic system. To study the microlymphatic endothelium, our laboratory isolated and cultured endothelial cells from microlymphatics. These cells were screened for endotheliam phenotype using a variety of endothelial- and lymphatic-specific markers. The defined RMLEC were passed and the profile of adhesion molecules expressed RMLEC was then determined using PCR, immunofluorescence, and Western blots. Further tests were done to determine the interaction of cadherins and catenins in RMLEC using immunoprecipitation studies. β-catenin was the only catenin which immunoprecipitated N-cadherin protein. β-catenin also was able to immunoprecipitate α-catenin and p120 protein. Variations in cadherin and catenin protein expression occurred in RMLEC with changes in passage number. VE-cadherin was undetected on RMLEC of passage number 15-20. Later studies on higher-passage cells showed that these cells expressed both VE-cadherin and N-cadherin, although the N-cadherin was present in larger amounts than VE-cadherin. Additionally, immunoprecipitation studies showed a decrease in N-cadherin and γ-catenin complexed with α-catenin in the older cells. The expression of mRNA and protein for key growth factors and their receptors was analyzed in RMLEC. Using immunocytochemistry and PCR, the three main isotypes of VEGF receptors, VEGFR-1, VEGFR-2 and VEGFR-3, were detected in RMLEC. RMLEC were also tested using immunofluorescence and PCR to look for ANGs and Tie receptor expression. RMLEC express mRNA for Ang-1, Ang-2, and Tie-2. In addition, all of these proteins except for Ang-1 were found on RMLEC by immunofluorescence. Development of the RMLEC cell line provides an appropriate in-vitro model for studies of microlymphatic endothelial function, such as lymphangiogenesis and endothelial cell modulation of the lymph pump. These results form the basis for future studies concerning which adhesion molecules are most important for cell-cell and cell-matrix adhesion in lymphatic endothelium, as well as what their involvement might be in normal function and pathological states. |
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| Item Description: | Vita. "Major Subject: Medical Sciences". "Submitted to the Graduate School of Biomedical Sciences of The Texas A&M University System Health Science Center in partial fulfillment for the requirements for the degree of Master of Science August 2005." Approved as to style and content by: Alan Parrish, David Zawieja, Emily Wilson, John Quarles. |
| Physical Description: | xii, 91 leaves : illustrations ; 28 cm. |
| Bibliography: | Includes bibliographical references (leaves 76-87). |