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Proceedings Paper

Investigation for the differentiation process of mouse ES cells by Raman spectroscopy
Author(s): Yoshinori Yamaguchi; Maha A. El-Hagrasy; Eiichi Shimizu; Masato Saito; Eiichi Tamiya
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Paper Abstract

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: 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.

Paper Details

Date Published: 9 February 2012
PDF: 8 pages
Proc. SPIE 8225, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X, 82250J (9 February 2012); doi: 10.1117/12.907586
Show Author Affiliations
Yoshinori Yamaguchi, Osaka Univ. (Japan)
Maha A. El-Hagrasy, Osaka Univ. (Japan)
Eiichi Shimizu, Osaka Univ. (Japan)
Masato Saito, Osaka Univ. (Japan)
Eiichi Tamiya, Osaka Univ. (Japan)

Published in SPIE Proceedings Vol. 8225:
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X
Daniel L. Farkas; Dan V. Nicolau; Robert C. Leif, Editor(s)

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