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D. Yuen, L. Chen; Novel Characterization of a Mouse Cell Line Expressing LYVE-1. Invest. Ophthalmol. Vis. Sci. 2009;50(13):1952.
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Lymphatic research represents a field of new discovery. The lymphatic pathway plays an essential role in corneal inflammation and transplantation immunity. The progress of lymphatic research is greatly hampered by the lack of mouse lymphatic endothelial cell lines for in vitro studies. Several widely used blood endothelial cell lines have been reported to be contaminated by lymphatic endothelial cells. The purpose of this study is to characterize a transformed mouse endothelial cell line, bEnd.3, and to study its potential as a model to investigate lymphangiogenesis in vitro.
Mouse bEnd.3 (ATCC, Manassas, VA) were cultured in EGM2 medium. The mRNA and protein expression of CD31 (a panendothelial cell marker), and several newly identified lymphatic specific markers including LYVE-1, Podoplanin, PROX1, and VEGFR-3 were investigated by reverse transcription PCR (RT-PCR), immunofluorescent microscopy, and flow cytometric analysis.
Surprisingly, our flow cytometric analysis showed that most of the bEnd.3 cells are CD31+LYVE+. Further investigations with both RT-PCR and immunofluorescent microscopy also confirmed their expression of CD31, LYVE-1, and VEGFR-3. However, these cells are all negative for PROX1 and podoplanin, two of the most commonly used lymphatic markers.
LYVE-1, a lymphatic marker, is expressed in the bEnd.3 cells. Further studies are undertaken to fully characterize and phenotype these cells.
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