RRC ID 61879
著者 Kamao H, Mandai M, Wakamiya S, Ishida J, Goto K, Ono T, Suda T, Takahashi M, Kiryu J.
タイトル Objective evaluation of the degree of pigmentation in human induced pluripotent stem cell-derived RPE.
ジャーナル Invest Ophthalmol Vis Sci
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.
巻・号 55(12)
ページ 8309-18
公開日 2014-11-11
DOI 10.1167/iovs.14-14694
PII iovs.14-14694
PMID 25389202
MeSH Embryonic Stem Cells / cytology* Embryonic Stem Cells / metabolism Epithelial Cells* / metabolism Eye Proteins / metabolism Humans Induced Pluripotent Stem Cells / cytology* Induced Pluripotent Stem Cells / metabolism Nerve Growth Factors / metabolism Observer Variation Pigmentation* Reproducibility of Results Retinal Pigment Epithelium / cytology* Retinal Pigment Epithelium / metabolism Serpins / metabolism Vascular Endothelial Growth Factor A / metabolism
IF 3.47
リソース情報
ヒト・動物細胞 253G1(HPS0002) 454E2(HPS0077)