Eye diseases such as age-related macular degeneration or diabetes affect retinal pigment epithelium (RPE) function and lead to retinal degeneration, vision loss and blindness. To study RPE function, physiology, and pathology, many laboratories have attempted to culture RPE as a more accessible alternative to native tissue. This goal has been accomplished with varying degrees of success due to the functional and morphological complexity of the RPE and its neighboring cells in the retina and the choroid. We have developed techniques for culturing confluent monolayers of human fetal RPE cells that exhibit morphology, physiology, and patterns of protein expression similar to native human fetal RPE. One of the goals of this work was to identify a set of commercially available ingredients to create stable and reproducible RPE cell cultures. We been able to produce confluent pigmented RPE cell cultures with classic epithelial morphology, transepithelial potential of 1 - 3mV, and transepithelial resistance greater than 400 Ohms*cm2. In the present experiments we further characterized these cultures using electron-microscopy and immunohistochemistry to identify cell structures, and localize apical and basolateral membrane and intercellular junctional complex proteins. ELISAs were used to confirm the polarity of secretion of selected cytokines. Intracellular microelectrodes were used to characterize receptor-mediated second messenger pathways and their downstream electrophysiological properties at the apical and basolateral membranes. The capacitance probe technique was used to measure net transepithelial fluid transport. This model has been used to help identify some of the mitochondrial pathways that mediate oxidative stress and the protective effects of lipoic acid. Gene signature of RPE was defined

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Investigator-Initiated Intramural Research Projects (ZIA)
Project #
1ZIAEY000419-06
Application #
7968352
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
6
Fiscal Year
2009
Total Cost
$444,057
Indirect Cost
Name
U.S. National Eye Institute
Department
Type
DUNS #
City
State
Country
Zip Code
Bulloj, Ayelen; Maminishkis, Arvydas; Mizui, Masayuki et al. (2017) Semaphorin4D-PlexinB1 Signaling Attenuates Photoreceptor Outer Segment Phagocytosis by Reducing Rac1 Activity of RPE Cells. Mol Neurobiol :
Benedicto, Ignacio; Lehmann, Guillermo L; Ginsberg, Michael et al. (2017) Concerted regulation of retinal pigment epithelium basement membrane and barrier function by angiocrine factors. Nat Commun 8:15374
Miyagishima, Kiyoharu J; Wan, Qin; Miller, Sheldon S et al. (2017) A basis for comparison: sensitive authentication of stem cell derived RPE using physiological responses of intact RPE monolayers. Stem Cell Transl Investig 4:
Miyagishima, Kiyoharu J; Wan, Qin; Corneo, Barbara et al. (2016) In Pursuit of Authenticity: Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium for Clinical Applications. Stem Cells Transl Med 5:1562-1574
Luna, Gabriel; Lewis, Geoffrey P; Linberg, Kenneth A et al. (2016) Anatomical and Gene Expression Changes in the Retinal Pigmented Epithelium Atrophy 1 (rpea1) Mouse: A Potential Model of Serous Retinal Detachment. Invest Ophthalmol Vis Sci 57:4641-54
Hotaling, Nathan A; Khristov, Vladimir; Wan, Qin et al. (2016) Nanofiber Scaffold-Based Tissue-Engineered Retinal Pigment Epithelium to Treat Degenerative Eye Diseases. J Ocul Pharmacol Ther 32:272-85
Blenkinsop, Timothy A; Saini, Janmeet S; Maminishkis, Arvydas et al. (2015) Human Adult Retinal Pigment Epithelial Stem Cell-Derived RPE Monolayers Exhibit Key Physiological Characteristics of Native Tissue. Invest Ophthalmol Vis Sci 56:7085-99
Shi, Guangpu; Chen, Siqi; Wandu, Wambui S et al. (2015) Inflammasomes Induced by 7-Ketocholesterol and Other Stimuli in RPE and in Bone Marrow-Derived Cells Differ Markedly in Their Production of IL-1? and IL-18. Invest Ophthalmol Vis Sci 56:1658-64
Ratnapriya, Rinki; Zhan, Xiaowei; Fariss, Robert N et al. (2014) Rare and common variants in extracellular matrix gene Fibrillin 2 (FBN2) are associated with macular degeneration. Hum Mol Genet 23:5827-37
Kay, Paul; Yang, Yit C; Hiscott, Paul et al. (2014) Age-related changes of cystatin C expression and polarized secretion by retinal pigment epithelium: potential age-related macular degeneration links. Invest Ophthalmol Vis Sci 55:926-34

Showing the most recent 10 out of 24 publications