Prof. Yoshinori Yamaguchi Profile

Prof. Yoshinori Yamaguchi

at Photonics Ctr Osaka Univ

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Proceedings Article | 9 February 2012 Paper

Investigation for the differentiation process of mouse ES cells by Raman spectroscopy

Yoshinori Yamaguchi, Maha El-Hagrasy, Eiichi Shimizu, Masato Saito, Eiichi Tamiya

Proceedings Volume 8225, 82250J (2012) https://doi.org/10.1117/12.907586

KEYWORDS: Raman spectroscopy, Proteins, Stem cells, Laser induced fluorescence, Calcium, Objectives, Tissues, Imaging spectroscopy, Raman scattering, In vitro testing

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The arrangement of differentiated pluripotent embryonic stem cells into three-dimensional aggregates, which are known as embryonic bodies, is a main step for progressing the embryonic stem cells differentiation. In this work, embryonic stem cells that were directly produced from the hanging drop step as a three-dimensional structure with no further twodimensional differentiation were diagnosed with Raman spectroscopy as a non-invasive and label-free technique. Raman spectroscopy was employed to discriminate between mouse embryonic bodies of different degrees of maturation. EBs were prepared applying the hanging drop method. The Raman scattering measurements were obtained in vitro with a Nanophoton RAMAN-11 micro-spectrometer (Japan: URL: www.nanophoton.jp equipped with an Olympus XLUM Plan FLN 20X/NA= 1.0 objective lens. Spectral data were smoothed, baseline corrected and normalized to the a welldefined intense 1003 cm-1 band (phenylalanine) which is insensitive to changes in conformation or environment. The differentiation process of embryonic stem cells is initiated by the removal of LIF from culture medium. 1, 7 and 17-dayold embryonic stem cells were collected and investigated by Raman spectroscopy. The main differences involve bands which decreased with maturation such as: 784 cm-1 (U, T, C ring br DNA/RNA, O-P-O str); 1177 cm-1 (cytosine, guanine) and 1578 cm-1 (G, A). It was found that with the progress of differentiation the protein content was amplified. The increase of protein to nucleic acid ratio was also previously observed with the progress of the differentiation process. Raman spectroscopy has the potential to distinguish between the Raman signatures of live embryonic stem cells with different degrees of maturation.

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