Basement membrane-dependent modification of phenotype and gene expression in human retinal pigment epithelial ARPE-19 cells
Abstract
PURPOSE: To use porcine lens capsule (PLC) as basement membrane for ARPE-19 cells and to characterize its effects on cell differentiation and gene expression. METHODS: Postconfluent cultures of ARPE-19 cells were established on either porous polyester filters or PLC membranes and characterized by electron microscopy, immunocytochemistry, and transepithelial electrical resistance measurements. Metabolic activity was assessed by measuring phagocytosis of rod outer segments. mRNA populations of ARPE-19 cells grown on polyester and PLC membranes were compared by suppressive subtractive hybridization. Differentially regulated messages were subsequently identified by DNA sequencing and their altered expression confirmed by Northern or virtual Northern blot analysis. RESULTS: Culture of ARPE-19 cells on PLC membrane induced the formation of apical microvilli and the ability to phagocytose rod outer segments. These culture conditions also led to enhanced junctional distribution of ZO-1 and occludin, the formation of polarized membrane domains, and a significant increase in transepithelial resistance. Gene expression was significantly altered by growth on PLC membranes and 29 differentially expressed transcripts were identified. CONCLUSIONS: Culture of ARPE-19 cells on PLC membranes resulted in a more differentiated phenotype and in expression of a specific set of transcripts encoding protein products that may affect epithelial differentiation, polarity and survival.
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Growth of ARPE-19 cells on extracellular matrix-rich membranes such as porcine lens capsule yields cells with improved phagocytosis and TER values reaching 200 Ω cm2.
“The second method of ARPE-19 cell differentiation is the growth of cells on extracellular matrix-rich membranes such as porcine lens capsule (Turowski et al., 2004). Such cells have been reported to have improved phagocytosis and TER values reaching 200 Ω cm2.”
From The cell biology of the retinal pigment epithelium (2020) · cites this paper as Turowski et al., 2004
ANXA2 is a membrane-associated protein that regulates actin dynamics and binds signaling intermediates, serving as a direct or indirect linker between the phagosome membrane and actin cytoskeleton.
“Annexin A2 (ANXA2) is a membrane-associated protein that regulates actin dynamics and binds signaling intermediates such as Ca2+, cholesterol-rich membranes, and phosphoinositides, serving as a direct or indirect linker between the phagosome membrane and actin cytoskeleton (Turowski et al., 2004; Law et al., 2009).”
From The cell biology of the retinal pigment epithelium (2020) · cites this paper as Turowski et al., 2004
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References 42 · Cited by 47
- Authors
- Turowski, P., Adamson, P., Sathia, J., Zhang, J. J., Moss, S. E., Aylward, G. W., Hayes, M. J., Kanuga, N., & Greenwood, J.
- Affiliations
- University College London (GB)
- Moorfields Eye Hospital NHS Foundation Trust (GB)
- Venue
- Investigative Opthalmology & Visual Science
- Published
- 2004-08-01
- Volume
- 45
- Issue
- 8
- Pages
- 2786
- ISSN
- 0146-0404 (ISSN-L)
- 1552-5783
- OpenAlex
- W2056674993
- Cited by
- 47
- References
- 42
- Citations by year
- 2021: 3 · 2020: 4 · 2019: 4 · 2018: 2 · 2017: 5 · 2014: 2 · 2013: 1 · 2012: 3
- FWCI
- 4.19
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Investigative Ophthalmology & Visual Science (publishedVersion)
- PubMed (publishedVersion)
- UCL Discovery (University College London) (submittedVersion)
- Topics
- Health Sciences › Medicine › Ophthalmology › Retinal Diseases and Treatments
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Retinal Development and Disorders
- Life Sciences › Neuroscience › Neurology › Barrier Structure and Function Studies
- Keywords
- retinal pigment epithelium, ARPE-19 cells, basement membrane, epithelial differentiation, cell polarity, ZO-1, occludin, tight junctions, transepithelial electrical resistance, rod outer segment phagocytosis, microvilli, gene expression, suppression subtractive hybridization, differentially expressed transcripts
- MeSH terms
- Animals, Basement Membrane, Cell Differentiation, Cell Line, Cell Survival, Electric Conductivity, Eye Proteins, Gene Expression Regulation, Humans, Immunohistochemistry, Lens Capsule, Crystalline, Membrane Proteins, Microscopy, Electron, Phagocytosis, Phenotype, Phosphoproteins, Pigment Epithelium of Eye, RNA, Messenger, Rod Cell Outer Segment, Swine, Blotting, Northern, Gene Library, Cell Polarity, Microscopy, Confocal, Cell Culture Techniques, Occludin, Zonula Occludens-1 Protein
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