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Title: Subretinal Pigment Epithelial Deposition of Drusen Components Including Hydroxyapatite in a Primary Cell Culture Model

Authors:
 [1];  [2];  [3];  [3];  [4];  [2];  [5];  [6];  [6];  [6];  [6]
  1. UCL Institute of Ophthalmology, University College London, London, United Kingdom 2Division of Biomaterials and Tissue Engineering, UCL Eastman Dental Institute, University College London, London, United Kingdom
  2. UCL Institute of Ophthalmology, University College London, London, United Kingdom 3Centre for Experimental Medicine, School of Medicine, Dentistry and Biomedical Science, Queen's University Belfast, Belfast, Northern Ireland, United Kingdom
  3. Center for Advanced Radiation Sources, The University of Chicago, Chicago, Illinois, United States
  4. Department of Materials, Imperial College London, London, United Kingdom
  5. Division of Biomaterials and Tissue Engineering, UCL Eastman Dental Institute, University College London, London, United Kingdom
  6. Department of Ophthalmology, University of Alabama School of Medicine, Birmingham, Alabama, United States
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
NIHFOREIGN
OSTI Identifier:
1343151
Resource Type:
Journal Article
Resource Relation:
Journal Name: Investigative Opthalmology & Visual Science; Journal Volume: 58; Journal Issue: 2
Country of Publication:
United States
Language:
ENGLISH

Citation Formats

Pilgrim, Matthew G., Lengyel, Imre, Lanzirotti, Antonio, Newville, Matt, Fearn, Sarah, Emri, Eszter, Knowles, Jonathan C., Messinger, Jeffrey D., Read, Russell W., Guidry, Clyde, and Curcio, Christine A.. Subretinal Pigment Epithelial Deposition of Drusen Components Including Hydroxyapatite in a Primary Cell Culture Model. United States: N. p., 2017. Web. doi:10.1167/iovs.16-21060.
Pilgrim, Matthew G., Lengyel, Imre, Lanzirotti, Antonio, Newville, Matt, Fearn, Sarah, Emri, Eszter, Knowles, Jonathan C., Messinger, Jeffrey D., Read, Russell W., Guidry, Clyde, & Curcio, Christine A.. Subretinal Pigment Epithelial Deposition of Drusen Components Including Hydroxyapatite in a Primary Cell Culture Model. United States. doi:10.1167/iovs.16-21060.
Pilgrim, Matthew G., Lengyel, Imre, Lanzirotti, Antonio, Newville, Matt, Fearn, Sarah, Emri, Eszter, Knowles, Jonathan C., Messinger, Jeffrey D., Read, Russell W., Guidry, Clyde, and Curcio, Christine A.. Wed . "Subretinal Pigment Epithelial Deposition of Drusen Components Including Hydroxyapatite in a Primary Cell Culture Model". United States. doi:10.1167/iovs.16-21060.
@article{osti_1343151,
title = {Subretinal Pigment Epithelial Deposition of Drusen Components Including Hydroxyapatite in a Primary Cell Culture Model},
author = {Pilgrim, Matthew G. and Lengyel, Imre and Lanzirotti, Antonio and Newville, Matt and Fearn, Sarah and Emri, Eszter and Knowles, Jonathan C. and Messinger, Jeffrey D. and Read, Russell W. and Guidry, Clyde and Curcio, Christine A.},
abstractNote = {},
doi = {10.1167/iovs.16-21060},
journal = {Investigative Opthalmology & Visual Science},
number = 2,
volume = 58,
place = {United States},
year = {Wed Feb 01 00:00:00 EST 2017},
month = {Wed Feb 01 00:00:00 EST 2017}
}
  • Extracellular deposits containing hydroxyapatite, lipids, proteins, and trace metals that form between the basal lamina of the RPE and the inner collagenous layer of Bruch’s membrane are hallmarks of early AMD. We examined whether cultured RPE cells could produce extracellular deposits containing all of these molecular components. Retinal pigment epithelium cells isolated from freshly enucleated porcine eyes were cultured on Transwell membranes for up to 6 months. Deposit composition and structure were characterized using light, fluorescence, and electron microscopy; synchrotron x-ray diffraction and x-ray fluorescence; secondary ion mass spectroscopy; and immunohistochemistry. Apparently functional primary RPE cells, when cultured on 10-lm-thickmore » inserts with 0.4-lm-diameter pores, can produce sub-RPE deposits that contain hydroxyapatite, lipids, proteins, and trace elements, without outer segment supplementation, by 12 weeks. In conclusion, the data suggest that sub-RPE deposit formation is initiated, and probably regulated, by the RPE, as well as the loss of permeability of the Bruch’s membrane and choriocapillaris complex associated with age and early AMD. This cell culture model of early AMD lesions provides a novel system for testing new therapeutic interventions against sub-RPE deposit formation, an event occurring well in advance of the onset of vision loss.« less
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  • A 62-year-old white female was found to have a small, flat pigmented choroidal tumor. After fluorescein angiography and a positive P-32 test, the eye was enucleated for presumed malignant melanoma. Histologically, the tumor proved to be a choroidal nevus with a break in Bruch's membrane and a subretinal-pigment-epithelial neovascular membrane.
  • Highlights: ► We established a 3D culture system to allow long-term culture of stomach cells. ► In this culture system, gastric epithelial cells grew for about 3 months. ► The cultured cells differentiated into multi-units of the stomach. ► This culture method should be useful for elucidating the cause of gastric diseases. -- Abstract: Compared to the small intestine and colon, little is known about stem cells in the stomach because of a lack of specific stem cell markers and an in vitro system that allows long-term culture. Here we describe a long-term three-dimensional (3D) primary gastric culture system withinmore » the stem cell niche. Glandular stomach cells from neonatal mice cultured in collagen gel yielded expanding sphere-like structures for 3 months. The wall of the gastrospheres consisted of a highly polarized epithelial monolayer with an outer lining of myofibroblasts. The epithelial cells showed a tall columnar cell shape, basal round nuclei, and mucus-filled cytoplasm as well as expression of MUC5AC, indicating differentiation into gastric surface mucous cells. These cells demonstrated the features of fully differentiated gastric surface mucous cells such as microvilli, junctional complexes, and glycogen and secretory granules. Fewer than 1% of cultured epithelial cells differentiated into enteroendocrine cells. Active proliferation of the epithelial cells and many apoptotic cells in the inner lumen revealed the rapid cell turnover in gastrospheres in vitro. This method enables us to investigate the role of signaling between cell–cell and epithelial–mesenchymal interactions in an environment that is extremely similar to the in vivo environment.« less