Parametric analysis of colony morphology of non-labelled live human pluripotent stem cells for cell quality control.
Given the difficulties inherent in maintaining human pluripotent stem cells (hPSCs) in a healthy state, hPSCs should be routinely characterized using several established standard criteria during expansion for research or therapeutic purposes. hPSC colony morphology is typically considered an important criterion, but it is not evaluated quantitatively. Thus, we designed an unbiased method to evaluate hPSC colony morphology. This method involves a combination of automated non-labelled live-cell imaging and the implementation of morphological colony analysis algorithms with multiple parameters. To validate the utility of the quantitative evaluation method, a parent cell line exhibiting typical embryonic stem cell (ESC)-like morphology and an aberrant hPSC subclone demonstrating unusual colony morphology were used as models. According to statistical colony classification based on morphological parameters, colonies containing readily discernible areas of differentiation constituted a major classification cluster and were distinguishable from typical ESC-like colonies; similar results were obtained via classification based on global gene expression profiles. Thus, the morphological features of hPSC colonies are closely associated with cellular characteristics. Our quantitative evaluation method provides a biological definition of 'hPSC colony morphology', permits the non-invasive monitoring of hPSC conditions and is particularly useful for detecting variations in hPSC heterogeneity.
Department of Basic Medicinal Sciences, Graduate School of Pharmaceutical Sciences, Nagoya University, Furocho, Chikusa-ku, Nagoya 464-8601, Japan.|Stem Cell Evaluation Technology Research Center (SCETRA), Hacho-bori, Chuou-ku, Tokyo 104-0032, Japan.
Ryuji
R
Kato
Department of Biotechnology, Graduate School of Engineering, Nagoya University, Furocho, Chikusa-ku, Nagoya 464-8602, Japan.
Megumi
M
Matsumoto
Department of Biotechnology, Graduate School of Engineering, Nagoya University, Furocho, Chikusa-ku, Nagoya 464-8602, Japan.
Hiroto
H
Sasaki
Department of Basic Medicinal Sciences, Graduate School of Pharmaceutical Sciences, Nagoya University, Furocho, Chikusa-ku, Nagoya 464-8601, Japan.
Risako
R
Joto
Department of Basic Medicinal Sciences, Graduate School of Pharmaceutical Sciences, Nagoya University, Furocho, Chikusa-ku, Nagoya 464-8601, Japan.
Mai
M
Okada
Department of Basic Medicinal Sciences, Graduate School of Pharmaceutical Sciences, Nagoya University, Furocho, Chikusa-ku, Nagoya 464-8601, Japan.
Yurika
Y
Ikeda
Department of Basic Medicinal Sciences, Graduate School of Pharmaceutical Sciences, Nagoya University, Furocho, Chikusa-ku, Nagoya 464-8601, Japan.
Kei
K
Kanie
Laboratory of Stem Cell Cultures, National Institutes of Biomedical Innovation, Health and Nutrition, Ibaraki, Osaka 567-0085, Japan.
Mika
M
Suga
Laboratory of Stem Cell Cultures, National Institutes of Biomedical Innovation, Health and Nutrition, Ibaraki, Osaka 567-0085, Japan.|Department of Cellular and Molecular Biology, Basic Life Sciences, Institute of Biomedical &Health Sciences, Hiroshima University, Hiroshima 734-8553, Japan.
Masaki
M
Kinehara
Laboratory of Stem Cell Cultures, National Institutes of Biomedical Innovation, Health and Nutrition, Ibaraki, Osaka 567-0085, Japan.
Kana
K
Yanagihara
Laboratory of Stem Cell Cultures, National Institutes of Biomedical Innovation, Health and Nutrition, Ibaraki, Osaka 567-0085, Japan.
Yujung
Y
Liu
Laboratory of Animal Models for Human Diseases, National Institutes of Biomedical Innovation, Health and Nutrition, Ibaraki, Osaka 567-0085, Japan.
Kozue
K
Uchio-Yamada
Laboratory of Stem Cell Cultures, National Institutes of Biomedical Innovation, Health and Nutrition, Ibaraki, Osaka 567-0085, Japan.
Takayuki
T
Fukuda
Stem Cell Evaluation Technology Research Center (SCETRA), Hacho-bori, Chuou-ku, Tokyo 104-0032, Japan.|Nikon Corporation, Nagaodaicho, Yokohama, Kanagawa 244-8533, Japan.
Hiroaki
H
Kii
Stem Cell Evaluation Technology Research Center (SCETRA), Hacho-bori, Chuou-ku, Tokyo 104-0032, Japan.|Nikon Corporation, Nagaodaicho, Yokohama, Kanagawa 244-8533, Japan.
Takayuki
T
Uozumi
Department of Biotechnology, Graduate School of Engineering, Nagoya University, Furocho, Chikusa-ku, Nagoya 464-8602, Japan.
Hiroyuki
H
Honda
Stem Cell Evaluation Technology Research Center (SCETRA), Hacho-bori, Chuou-ku, Tokyo 104-0032, Japan.|Nikon Corporation, Nagaodaicho, Yokohama, Kanagawa 244-8533, Japan.
Yasujiro
Y
Kiyota
Stem Cell Evaluation Technology Research Center (SCETRA), Hacho-bori, Chuou-ku, Tokyo 104-0032, Japan.|Laboratory of Stem Cell Cultures, National Institutes of Biomedical Innovation, Health and Nutrition, Ibaraki, Osaka 567-0085, Japan.
Miho K
MK
Furue
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