Abstract:
PURPOSE:Genetic modification of cells before transplantation may allow the delivery of neuroprotective and other functional molecules to patients with neurodegenerative diseases. To avoid complications associated with virally transfected cells, we have explored the use of nonviral methods to insert genetic material into RPE cells. METHODS:After transfection with plasmids encoding different pigment epithelium-derived factor (PEDF) fusion proteins, transfected cells were established and passaged up to 100 times. Gene expression of PEDF, ZnT3, ZIP2, CRALBP, CATD, and ZO-1 was determined by RT-PCR. Secretion dynamics were analyzed using ELISA and a spheroid-based assay was used to confirm the anti-angiogenic activity of the recombinant PEDF. RESULTS:Transfection efficiency reached up to 98.7% with a plasmid encoding PEDF and enhanced green fluorescent protein (EGFP) separately and 87.2% with a plasmid encoding an EGFP-PEDF fusion. Immunoblotting revealed that transfected RPE cells express the appropriate PEDF or EGFP-PEDF. Expression of recombinant PEDF is stable, as shown by its secretion for the 2 years and the 100 passages the cells have been followed. PEDF expression was overexpressed and the transfected cells exhibited increased proliferation, up-regulation of ZnT3 and ZIP2, and inhibited sprouting in human umbilical vein endothelial cell spheroids. CONCLUSIONS:Genetic in vitro modification of pigment epithelial cells using nonviral transfection protocols should improve the potential therapeutic treatment of neurodegenerative diseases by transplantation of genetically modified cells without the disadvantages of virally mediated transfection. Here we have shown that genetically modified RPE cells overexpress a functional human recombinant PEDF, as evidenced by the autogenic regulation of proliferation, up-regulation of two distinct zinc transporters, and in vitro inhibition of endothelial cell sprouting.
journal_name
Invest Ophthalmol Vis Scijournal_title
Investigative ophthalmology & visual scienceauthors
Johnen S,Kazanskaya O,Armogan N,Stickelmann C,Stöcker M,Walter P,Thumann Gdoi
10.1167/iovs.10-6178subject
Has Abstractpub_date
2011-07-23 00:00:00pages
5400-7issue
8eissn
0146-0404issn
1552-5783pii
iovs.10-6178journal_volume
52pub_type
杂志文章abstract:PURPOSE:Lectin studies have shown that in the rabbit corneal epithelium, alpha-2,3 sialylation of O-linked glycans differentiates limbal and corneal epithelial cell phenotypes. Because sialic acid can be regulated at the level of the expression of sialyltransferases (STs), the purpose of the present study was to analyz...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1998-05-01 00:00:00
abstract:PURPOSE:The goal of this study was to determine the changes in the organization of the retinal cytoskeleton after experimental retinal detachment. METHODS:Cat retinas were detached from the retinal pigment epithelium and then processed for Western blot analysis and fluorescence microscopy. Proteins examined included g...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1995-11-01 00:00:00
abstract:PURPOSE:To examine the cross-sectional relationship between blood pressure, perfusion pressure, and prevalence of open angle glaucoma (OAG) in an adult Latino population. METHODS:Participants aged 40 years and older (N = 6130) from the Los Angeles Latino Eye Study (LALES), a large, population-based study of self-ident...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.08-2956
更新日期:2010-06-01 00:00:00
abstract:PURPOSE:To study different aspects of macular function in eyes with early age-related macular degeneration (early AMD: drusen with or without retinal pigment epithelium alterations) and normal visual acuity, to obtain a complete evaluation of macular function impairment in early AMD and to study the relationship betwee...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1997-02-01 00:00:00
abstract::Eight murine monoclonal antibodies to retinal S-antigen were found to recognize three antigenic clusters in competitive binding experiments. Variable patterns of immunohistochemical staining of retinal sections supported the competitive binding studies. Monoclonal antibodies to two of these three antigenic clusters on...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1988-01-01 00:00:00
abstract:PURPOSE:To evaluate the performance of an automated retinal image quality assessment system for use in automated diabetic retinopathy grading. METHODS:Algorithmic methods have been developed for assessing the quality of 45 degrees single field retinal images for use in diabetic retinopathy screening. For this purpose,...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.05-1155
更新日期:2006-03-01 00:00:00
abstract:Purpose:To refine the Minnesota Grading System (MGS) using definitions from the Age-Related Eye Disease Studies (AREDS) into a nine-step grading scale (MGS-9). Methods:A nine-step grading scale descriptive analysis using three key phenotypic features (total drusen area, increased, and decreased pigmentation) of human ...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.17-22161
更新日期:2017-10-01 00:00:00
abstract:PURPOSE:To characterize photoreceptor cell apoptosis and cell loss in a mouse model of experimental retinal detachment (RD), and to use the technology of mouse genetics to study the molecular mechanisms underlying RD-associated photoreceptor degeneration. METHODS:Retinal detachments were created in adult wild-type and...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.03-0827
更新日期:2004-02-01 00:00:00
abstract:PURPOSE:Adipogenesis contributes to the pathogenesis of thyroid eye disease (TED). Thyrotropin receptor (TSHR) transcripts are present in orbital fat. This study was conducted to determine whether they are expressed as functional protein, and if so, whether this is restricted to TED orbits or to a particular stage in a...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:2000-10-01 00:00:00
abstract:PURPOSE:To investigate the effect of varying levels of ocular surface stimulation on the timing and amplitude of the blink and tear secretion. METHODS:Following instillation of fluorescein dye, increasing levels of air flow were directed toward the central corneas of 10 healthy subjects. Interblink interval (IBI), tea...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.14-16313
更新日期:2015-07-01 00:00:00
abstract:PURPOSE:The use of patient-reported outcome (PRO) measures to assess self-reported difficulty in visual activities is common in patients with impaired vision. This study determines the visual and psychosocial factors influencing patients' responses to self-report measures, to aid in understanding what is being measured...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.10-7055
更新日期:2011-07-15 00:00:00
abstract:PURPOSE:Achieving good vision in infants born with a unilateral cataract is believed to require early surgery and consistent occlusion of the fellow eye. This article examines the relationship between adherence to patching and grating acuity. METHODS:Data came from the Infant Aphakia Treatment Study, a randomized clin...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章,多中心研究,随机对照试验
doi:10.1167/iovs.11-8457
更新日期:2012-06-05 00:00:00
abstract:PURPOSE:A great deal of information about functionally significant domains of a protein may be obtained by comparison of primary sequences of gene homologues over a broad phylogenetic base. This study was designed to identify evolutionarily conserved domains of the photoreceptor disc membrane protein peripherin/rds by ...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.02-1152
更新日期:2003-06-01 00:00:00
abstract::Visual fixation of a small, stationary target was measured in 12 young observers (mean age = 22 yr) and in 12 older observers (mean age = 70 yr). The two groups' fixation behavior did not differ on various dimensions of fixation stability: mean fixation area, intra-subject variability, or changes in fixation over succ...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1986-12-01 00:00:00
abstract:PURPOSE:To investigate whether time outdoors and a range of other activities are associated with change in spherical equivalent (SE) and axial length in Chinese children over a period of 2 years. METHODS:A total of 1997 children aged 12.7 ± 0.5 (10.9-15.6) years in the Anyang Childhood Eye Study (ACES) were examined a...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.14-15474
更新日期:2015-07-01 00:00:00
abstract:PURPOSE:To study retinal structure in choroideremia patients and carriers using high-resolution imaging techniques. METHODS:Subjects from four families (six female carriers and five affected males) with choroideremia (CHM) were characterized with best-corrected visual acuity (BCVA), kinetic and static perimetry, full-...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.12-10707
更新日期:2013-02-01 00:00:00
abstract:PURPOSE:To evaluate the feasibility of near-infrared (NIR) imaging acquisition in a large sample of consecutive pediatric patients with neurofibromatosis type 1 (NF1), to evaluate the diagnostic performance of NF1-related choroidal abnormalities as a diagnostic criterion of the disease, and to compare this criterion wi...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.14-16053
更新日期:2015-09-01 00:00:00
abstract::The authors have examined the fate of intravitreally injected liposomes in the aphakic, vitrectomized eye of the rabbit. Liposomes labelled with 125[I]-p-hydroxybenzimidylphosphatidylethanolamine were eliminated rapidly from the intraocular fluid. Nonetheless, a significant fraction of these liposomes were found to bi...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1987-05-01 00:00:00
abstract:Purpose:Rose bengal (RB)-photosensitized protein crosslinking has been proposed for several applications in the eye. This study identifies oxygen-dependent and oxygen-independent mechanistic pathways in cornea for RB-photosensitized crosslinking to enhance its efficiency for ocular treatments. Methods:Rabbit corneas e...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.19-26604
更新日期:2019-05-01 00:00:00
abstract:PURPOSE:Intrastromal inlays for refractive correction of presbyopia are being adopted into clinical practice. An important concern is the effect of the inlay on the long-term health of the cornea due to disturbances in the concentration profiles of metabolic species. A three-dimensional metabolic model for the cornea i...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
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更新日期:2014-05-15 00:00:00
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journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.10-5438
更新日期:2010-09-01 00:00:00
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journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1978-09-01 00:00:00
abstract:PURPOSE:Experimental autoimmune uveoretinitis (EAU) in mice is an important model for elucidating basic mechanisms in autoimmune eye disease. The need for pertussis toxin (PTX) as an additional adjuvant to elicit EAU has limited the usefulness of this model in some types of studies by introducing a pleiotropic factor w...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1999-11-01 00:00:00
abstract::Parallel studies of the ionic balance and membrane permeability characteristics of normal human lenses were carried out in three countries (USA, England and Italy). Similar age-related changes were found in each laboratory. The lens membrane potential and resistance declined markedly with age while internal Na+ and fr...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1989-08-01 00:00:00
abstract::In the retina from an eye enucleated for choroidal melanoma apposed rod outer segments were seen by transmission electron microscopy to be fusing together in groups by the confluence of their plasma membranes and the formation of cytoplasmic bridges between adjacent outer segments. These fusions were seen in areas dis...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1977-07-01 00:00:00
abstract:Purpose:The purpose of this study was to study the status and association among tear-soluble factors, corneal dendritic cell density, vitamin D, and signs and symptoms in dry eye disease (DED). Methods:A total of 33 control subjects and 47 evaporative dry eye patients were included in the study. DED diagnosis and clas...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.19-26914
更新日期:2019-06-03 00:00:00
abstract:PURPOSE:To determine directional changes in peripheral corneal thickness and corneal astigmatism over a range of severity of Fuchs' endothelial corneal dystrophy (FECD). METHODS:Eyes with FECD were categorized as mild, moderate, or advanced according to the area and confluence of guttae and the presence of clinical ed...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.15-17311
更新日期:2015-09-01 00:00:00
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journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.12-11268
更新日期:2013-07-10 00:00:00
abstract:PURPOSE:Identification of causative mutations for retinal blinding disorders is often limited by restricted understanding of gene expression and underlying molecular mechanisms that trigger degenerative processes. This study was conducted to develop a catalog of canine retina-expressed genes that would provide a unique...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:10.1167/iovs.05-1463
更新日期:2006-06-01 00:00:00
abstract::The first heritable model of retinoblastoma was established by retina-specific expression of simian virus 40 T-antigen (SV40 T-ag) in transgenic mice. Bilateral, multifocal ocular tumors were observed in 100% of transgene-bearing mice. Central nervous system neoplasms occurred at a lower rate (27%) and represented the...
journal_title:Investigative ophthalmology & visual science
pub_type: 杂志文章
doi:
更新日期:1991-02-01 00:00:00