Analysis of Retinal Pigment Epithelium (RPE) in Human Eyes Open Access

Arora, Shagun Kumari (2014)

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Purpose: Describe RPE morphometry of normal human eyes with regard to age and topographic location. Methods: Cadaveric human donor eyes of varying ages were microdissected, RPE flatmounts were immunostained for AF635-phalloidin, nuclei stained by propidium iodide, and imaged by confocal miscroscopy. Image analysis was performed using ImageJ (NIH) and Cellprofiler software (Broad Institute, Cambridge, MA). A large number of quantitative parameters, including cell density, cell area, percentage of hexagonal cells, and measures of cell shape, were obtained from these analyses to characterize individual and groups of RPE cells. Measurements were taken from selected areas spanning the length of the retina, from the macula, mid-periphery, and far-periphery. Results: Fifteen eyes from 10 donors of varying ages ranging from 29-80 years were used. In the macula and far-periphery, an overall trend of decreasing RPE cell density, percent hexagonal cells and form factor was observed with increasing age. We also found a trend of increasing cell area and eccentricity with age in the macula and far-periphery. When individuals were divided into two age groups, <60 years old and 60 years old, there was a higher cell density, lower cell area, lower eccentricity and higher form factor in the younger group (p< 0.05 for all measurements). No significant differences in RPE morphometry between age groups were found in the mid-periphery. Conclusions: RPE cells become less dense, larger in size, lose their typical hexagonal shape and become more oval in shape with age.

Table of Contents

Table of Contents







Tables and Figures

Table 1: Summary of Past Studies...23

Figure 1: Study Population Diagram...25

Table 2: Donor Characteristics...26

Figure 2: Description of Methods...27

Figure 3: Image Selection...28

Figure 4: Image Analysis Technique...29

Table 3: Descriptive Statistics of RPE Morphometric Measurements...30

Figure 5: Correlation Analysis...31

Figure 6: Analysis of Spatial Variation of RPE Morphometry...32

Figure 7: Age-Related Morphometric Changes of RPE in 3 Areas of Retina...33
Table 4: Average Morphometric Measurements Comparing
Patients <60 Years and ≥60 Years of Age...34

Figure 8: Representative Images of RPE Flatmounts...35

Table 5: Linear Regression Model Parameter Estimates...36

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