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Comprehensive analysis of HEK293 cells reveals a LEC-like phenotype

Phillips, Nicole (2014) Comprehensive analysis of HEK293 cells reveals a LEC-like phenotype. Master's Thesis, University of Pittsburgh. (Unpublished)

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The history of cell culture dates back to 1907 when Ross Harrison discovered that neuronal cells could be cultured in vitro. Two types of cells used within cell culture include primary and established cell lines. Among established cells HEK 293 and 293T cells are among the most widely used in the technique of cell culture. Lymphatic endothelial cells (LECs) are often represented as primary cells, which have been isolated from tissues. Previous studies have characterized 293 and 293T cells as neuronal in origin although phenotypic analyses have not been reported. Similarly, studies dealing with the binding characteristics of HIV have revealed the expression of podoplanin, a typical LEC marker, by 293T cells. Additionally, our lab has observed the expression of podoplanin by 293 cells through flow cytometry analysis. These observations of podoplanin expression suggest that the phenotype of 293 and 293T cells needs to be further explored and that they might resemble LECs. This study outlines the comprehensive mRNA and protein analysis of 293 and 293T cells in the context of LECs using standard and real-time RT-PCR and flow cytometry analysis. Furthermore, this study evaluates the influence of culture conditions on LEC marker expression by 293 and 293T cells in an attempt to find optimal LEC growth conditions for these cell lines. Evaluation of mRNA expression levels revealed that 293 and 293T cells express multiple LEC markers including Prox-1, Lyve-1, PDPN, and VEGFR-3. Studies investigating the manipulation of culture conditions revealed that 293 and 293T cells do not significantly change in LEC marker expression. Taken together, these studies suggest that 293 and 293T cells phenotypically resemble LECs and should be defined as LEC-like. Defining 293 and 293T cells as LECs is highly relevant to the field of public health as it provides a new model by which to study LEC function in several different contexts including vaccine interaction and cancer metastasis.


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Item Type: University of Pittsburgh ETD
Status: Unpublished
CreatorsEmailPitt UsernameORCID
Phillips, Nicolenlp25@pitt.eduNLP25
ETD Committee:
TitleMemberEmail AddressPitt UsernameORCID
Thesis AdvisorReinhart, Todd A.reinhar@pitt.eduREINHAR
Committee MemberMarques, Ernesto T Amarques@pitt.eduMARQUES
Committee MemberDavidson, Lance Alad43@pitt.eduLAD43
Date: 27 June 2014
Date Type: Publication
Defense Date: 17 April 2014
Approval Date: 27 June 2014
Submission Date: 7 April 2014
Access Restriction: 2 year -- Restrict access to University of Pittsburgh for a period of 2 years.
Number of Pages: 90
Institution: University of Pittsburgh
Schools and Programs: School of Public Health > Infectious Diseases and Microbiology
Degree: MS - Master of Science
Thesis Type: Master's Thesis
Refereed: Yes
Uncontrolled Keywords: HEK293, LEC-like, 293, 293T, HDLEC, SV40,
Date Deposited: 27 Jun 2014 21:47
Last Modified: 15 Nov 2016 14:18


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