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Culturing of Retinal Pigment Epithelial Cells on an Ex Vivo Model of Aged Human Bruch's Membrane
Published on: April 12, 2018
Comparison of FRPE and human embryonic stem cell-derived RPE behavior on aged human Bruch's membrane
Ilene K Sugino1, Qian Sun, Jianqiu Wang
1The Institute of Ophthalmology and Visual Science, New Jersey Medical School, University of Medicine and Dentistry of New Jersey, 90 Bergen Street, Newark, NJ 07101, USA.
Investigative Ophthalmology & Visual Science
|April 5, 2011
Summary
Human embryonic stem cell-derived retinal pigment epithelium (hES-RPE) and fetal RPE (fRPE) show different behaviors on Bruch's membrane, suggesting hES-RPE may not fully replicate native RPE function in age-related macular degeneration (AMD) contexts.
Area of Science:
- Ophthalmology
- Stem Cell Biology
- Regenerative Medicine
Background:
- The retinal pigment epithelium (RPE) is crucial for retinal health, and its dysfunction is implicated in age-related macular degeneration (AMD).
- Human embryonic stem cell-derived RPE (hES-RPE) offers a potential alternative to native RPE for therapeutic applications.
Purpose of the Study:
- To compare the behavior of hES-RPE and fetal RPE (fRPE) when cultured on human Bruch's membrane (BM) from aged and AMD donors.
- To investigate differences in attachment, proliferation, marker retention, and protein secretion between hES-RPE and fRPE on BM.
Main Methods:
- Culturing hES-RPE (3 pigmentation levels) and fRPE on human BM explants.
- Assessing cell behavior using light, confocal, and scanning electron microscopy.
- Analyzing integrin mRNA levels via real-time PCR and secreted proteins using multiplex analysis.
Main Results:
- hES-RPE exhibited impaired initial attachment and limited BM resurfacing compared to fRPE.
- Pigmented hES-RPE showed similar nuclear densities to fRPE at day 21, but earlier time points revealed dissimilar cell behavior, proliferation, and marker retention.
- Protein secretion profiles differed, with hES-RPE secreting more nerve growth factor and TSP-2, while fRPE secreted more VEGF and BDNF on submacular BM.
Conclusions:
- Despite similar limited BM resurfacing by day 21, hES-RPE and fRPE displayed distinct behaviors in earlier culture stages.
- Differences in secreted proteins suggest that hES-RPE may not perfectly mimic native RPE function when transplanted onto aged or AMD-affected BM.
