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Comparative analysis of two ocular endothelial cells
Author(s) -
MAMMADZADA P,
GUDMUNDSSON J,
KVANTA A,
ANDRE H
Publication year - 2014
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2014.t067.x
Subject(s) - cd31 , immunostaining , choroid , endothelial stem cell , staining , angiogenesis , retina , cell type , cell , cell culture , endothelium , pathology , biology , microbiology and biotechnology , chemistry , in vitro , medicine , immunohistochemistry , cancer research , biochemistry , genetics , neuroscience
Purpose In the eye, angiogenesis is associated with many pathological conditions. Retinal and choroidal endothelial cells (RE and CE cells) are the main cellular components of ocular angiogenesis. It is widely accepted that endothelial cells (EC) from different vascular beds show heterogeneity. Comparative study of RE and CE cells can lead to deeper understanding of the mechanisms through which EC become activated to undergo pathologic behaviors. Therefore, the goal of the study was isolation, cell culturing, and characterization of human CE and RE cells, and comparative analysis of these two cell lines. Methods Human consented‐donor eyes were used for preparation of CE and RE cell cultures. The posterior segment of these eyes was microdissected into isolated tissues (retina and choroid). Isolation of RE and CE cells from these tissues was achieved by positive selection of endothelium by targeting CD31. Characterization of RE and CE cells was performed by immunostaining for EC specific markers (CD31, isolectin, and acetylated‐LDL uptake). Further analysis is achieved by qPCR. Results Endothelial cells from the retina and the choroid have been specifically isolated and displayed high endothelial cell‐type purity. Both RE and CE cell cultures displayed positive staining for CD31 and isolectin, as well as acetylated‐LDL uptake; positive staining was confirmed to 100% of the cultured cells. Differences between RE and CE cells was not clear upon IF microscopy. Initial qPCR analysis has shown discreet variations between RE and CE cells. Conclusion RE and CE cells exhibit similar morphologies and staining patterns for the specific endothelial cell markers studied. Extensive comparative analysis of these two ocular EC‐lines is undergoing by a series of qPCR arrays.