Embryonic stem cell differentiation studied by FT-IR spectroscopy

被引:97
作者
Ami, Diletta [1 ]
Neri, Tui [2 ]
Natalello, Antonino [1 ]
Mereghetti, Paolo [3 ]
Doglia, Silvia Maria [1 ]
Zanoni, Mario [2 ]
Zuccotti, Maurizio [4 ]
Garagna, Silvia [2 ]
Redi, Carlo Alberto [2 ,5 ]
机构
[1] Univ Milano Bicocca, Dipartimento Biotecnol & Biosci, I-20126 Milan, Italy
[2] Univ Stud Pavia, Dipartimento Biol Anim, Lab Biol Sviluppo, I-27100 Pavia, Italy
[3] Univ Sassari, Dipartimento Chim, I-07100 Sassari, Italy
[4] Univ Stud Parma, Sezione Istol Embriol, Dipartimento Med Sperimentale, I-43100 Parma, Italy
[5] Fondazione IRCCS Policlin San Matteo Pavia, Pavia, Italy
来源
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH | 2008年 / 1783卷 / 01期
关键词
cardiomyocyte; cytodifferentiation; embryonic stem cell; Fourier transform infrared spectroscopy; linear discriminant analysis; principal component analysis;
D O I
10.1016/j.bbamcr.2007.08.003
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We propose, here, an FT-IR method to monitor the spontaneous differentiation of murine embryonic stem (ES) cells in their early development. Principal component analysis and subsequent linear discriminant analysis enabled us to segregate stem cell spectra into separate clusters corresponding to different differentiation times - and to identify the most significant spectral changes during differentiation. Between days 4 to 7 of differentiation, these spectral changes in the protein amide I band (1700-1600 cm(-1)) and in the nucleic acid absorption region (1050-850 cm(-1)) indicated that mRNA translation was taking place and that specific proteins were produced, reflecting the appearance of a new phenotype. The DNA/ RNA hybrid bands (954 cm(-1) and 899 cm(-1)) were also observed, suggesting that the transcriptional switch of the genome started at this stage of differentiation, As confirmed by cytochemical assays, the FT-IR approach presented here allows to detect at molecular level the biological events of ES cell differentiation as they take place and to monitor in a rapid way the temporal evolution of the ES cell culture. (C) 2007 Elsevier B.V. All rights reserved.
引用
收藏
页码:98 / 106
页数:9
相关论文
共 45 条
[1]   Kinetics of inclusion body formation studied in intact cells by FT-IR spectroscopy [J].
Ami, D ;
Natalello, A ;
Gatti-Lafranconi, P ;
Lotti, M ;
Doglia, SM .
FEBS LETTERS, 2005, 579 (16) :3433-3436
[2]   Fourier transform infrared micro spectroscopy as a new tool for nematode studies [J].
Ami, D ;
Natalello, A ;
Zullini, A ;
Doglia, SM .
FEBS LETTERS, 2004, 576 (03) :297-300
[3]   FT-IR study of heterologous protein expression in recombinant Escherichia coli strains [J].
Ami, D ;
Bonecchi, L ;
Calì, S ;
Orsini, G ;
Tonon, G ;
Doglia, SM .
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS, 2003, 1624 (1-3) :6-10
[4]   Structural analysis of protein inclusion bodies by Fourier transform infrared microspectroscopy [J].
Ami, Diletta ;
Natalello, Antonino ;
Taylor, Geoffrey ;
Tonon, Giancarlo ;
Doglia, Silvia Maria .
BIOCHIMICA ET BIOPHYSICA ACTA-PROTEINS AND PROTEOMICS, 2006, 1764 (04) :793-799
[5]   Structure and dynamics of membrane proteins as studied by infrared spectroscopy [J].
Arrondo, JLR ;
Goñi, FM .
PROGRESS IN BIOPHYSICS & MOLECULAR BIOLOGY, 1999, 72 (04) :367-405
[6]   EMBRYONIC STEM-CELLS EXPRESS NEURONAL PROPERTIES IN-VITRO [J].
BAIN, G ;
KITCHENS, D ;
YAO, M ;
HUETTNER, JE ;
GOTTLIEB, DI .
DEVELOPMENTAL BIOLOGY, 1995, 168 (02) :342-357
[7]   A library of IR bands of nucleic acids in solution [J].
Banyay, M ;
Sarkar, M ;
Gräslund, A .
BIOPHYSICAL CHEMISTRY, 2003, 104 (02) :477-488
[8]   What vibrations tell us about proteins [J].
Barth, A ;
Zscherp, C .
QUARTERLY REVIEWS OF BIOPHYSICS, 2002, 35 (04) :369-430
[9]  
Bentley AJ, 2007, MOL VIS, V13, P237
[10]   In situ characterization of beta-amyloid in Alzheimer's diseased tissue by synchrotron Fourier transform infrared Microspectroscopy [J].
Choo, LP ;
Wetzel, DL ;
Halliday, WC ;
Jackson, M ;
LeVine, SM ;
Mantsch, HH .
BIOPHYSICAL JOURNAL, 1996, 71 (04) :1672-1679