Hiroyuki Kamao1, Michiko Mandai2, Shunji Wakamiya3, Junko Ishida3, Katsutoshi Goto3, Takaaki Ono3, Taiji Suda4, Masayo Takahashi2, Junichi Kiryu3. 1. Department of Ophthalmology, Kawasaki Medical School, Kurashiki, Okayama, Japan Laboratory for Retinal Regeneration, RIKEN Center for Developmental Biology, Chuo-ku, Kobe, Hyogo, Japan. 2. Laboratory for Retinal Regeneration, RIKEN Center for Developmental Biology, Chuo-ku, Kobe, Hyogo, Japan. 3. Department of Ophthalmology, Kawasaki Medical School, Kurashiki, Okayama, Japan. 4. Tissue Biology & Electron Microscopy Research Center, Kawasaki Medical School, Kurashiki, Okayama, Japan.
Abstract
PURPOSE: For the transplantation of human induced pluripotent stem cell-derived retinal pigment epithelium (hiPSC-RPE), determination of the maturation status of these cells is essential, and the degree of pigmentation (dPG) can serve as a good indicator of this status. The aim of this study was to establish a method of objectively and quantitatively evaluating the dPG of hiPSC-RPE. METHODS: Two observers determined the dPG subjectively by observing recorded images of hiPSC-RPE as follows: the dPG of a single cell was classified into three different pigmentation stages, and the overall dPG was compared between two cell groups to identify the group with the higher dPG. The κ statistic was applied to assess interobserver reproducibility. Next, the dPG of single cells and cell groups was objectively determined by the lightness of the hue, saturation, and value (HSL) color space, and the correlation between the subjective evaluation and time-dependent change in the objective dPG of hiPSC-RPE was investigated. RESULTS: The κ statistic was 0.88 and 0.81 in the single-cell and cell-group observations, respectively. The objective dPG of single cells and cell groups was highly correlated with the subjective dPG. However, the observers were occasionally unable to subjectively determine the group with the higher dPG. The objective dPG increased in a time-dependent manner. CONCLUSIONS: The lightness of the HSL color space can be used to objectively and quantitatively evaluate the dPG of hiPSC-RPE in culture. The objective evaluation was consistent and was able to better identify small differences than subjective evaluation. Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.
PURPOSE: For the transplantation of human induced pluripotent stem cell-derived retinal pigment epithelium (hiPSC-RPE), determination of the maturation status of these cells is essential, and the degree of pigmentation (dPG) can serve as a good indicator of this status. The aim of this study was to establish a method of objectively and quantitatively evaluating the dPG of hiPSC-RPE. METHODS: Two observers determined the dPG subjectively by observing recorded images of hiPSC-RPE as follows: the dPG of a single cell was classified into three different pigmentation stages, and the overall dPG was compared between two cell groups to identify the group with the higher dPG. The κ statistic was applied to assess interobserver reproducibility. Next, the dPG of single cells and cell groups was objectively determined by the lightness of the hue, saturation, and value (HSL) color space, and the correlation between the subjective evaluation and time-dependent change in the objective dPG of hiPSC-RPE was investigated. RESULTS: The κ statistic was 0.88 and 0.81 in the single-cell and cell-group observations, respectively. The objective dPG of single cells and cell groups was highly correlated with the subjective dPG. However, the observers were occasionally unable to subjectively determine the group with the higher dPG. The objective dPG increased in a time-dependent manner. CONCLUSIONS: The lightness of the HSL color space can be used to objectively and quantitatively evaluate the dPG of hiPSC-RPE in culture. The objective evaluation was consistent and was able to better identify small differences than subjective evaluation. Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.
Entities:
Keywords:
degree of pigmentation; human induced pluripotent stem cell; retinal pigment epithelium
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